DE69418947T2 - STYLE DERIVATIVES AND ANALOGS AND THEIR USE - Google Patents

Description

The present invention relates to novel compounds of a stilbene derivative, a stilbene homolog derivative and an alpha,beta-diarylacrylonitrile derivative selectively inhibiting 12-lipoxygenase, and pharmaceutical compositions containing the foregoing compounds as effective ingredients, more particularly, the present invention relates to a stilbene derivative, a stilbene homolog derivative and an alpha,beta-diarylacrylonitrile derivative having a selective effect of inhibiting the activities of 12-lipoxygenase in the course of 12-lipoxygenase, and pharmaceutical compositions containing these compounds as effective ingredients and selectively inhibiting 12-lipoxygenase and usable as pharmaceutical compositions for preventing and curing various circulatory diseases such as arteriosclerosis and vascular spasms, and preventing the metastasis of some cancers (e.g. Lewis lung cancer).

Furthermore, the present invention relates to novel compounds of a stilbene derivative and a stilbene homologue derivative capable of producing a substance whose modified part is cleaved in vivo and which selectively inhibits 12-lipoxygenase; and pharmaceutical compositions containing the above-mentioned compounds as active ingredients; more particularly, the present invention relates to a stilbene derivative and a stilbene homologue derivative (these may be described as precursors hereinafter) capable of being converted into compounds whose modified parts are cleaved in vivo and which selectively inhibit lipoxygenase in the course of 12-lipoxygenase; and medicines containing these precursors as active ingredients and selectively inhibiting 12-lipoxygenase and usable as medicines for the prevention and cure of various circulatory diseases, such as arteriosclerosis and vascular spasms, as well as the prevention of metastasis of some types of cancer.

The percentages in this description are based on weight unless otherwise stated.

Technical background

It is known that there is a metabolic pathway in the arachidonate cascade called 5-lipoxygenase pathway and that arachidonate is converted to 5-hydroperoxyeicosatetraenoic acid by the function of 5-lipoxygenase ("Prostaglandin and Morbid States", edited by Seiitsu Murota, Tokyo Kagaku Dojin, 1984).

It is known that numerous leukotrienes are biosynthesized with this compound as an intermediate ("Prostaglandin and Morbid States", edited by Seiitsu Mourota, Tokyo Kagaku Dojin, 1984), and it is also known that, for example, among these leukotrienes, leukotriene B4 has a strong effect on leukocyte migration and is a mediator of inflammation, and that leukotriene C4 and D4 are mediators of asthma ("Prostaglandin and Morbid States", edited by Seiitsu Murota, Tokyo Kagaku Dojin, 1984).

Since then, an extensive search for drugs with the effect of 5-lipoxygenase inhibition has been carried out from the point of view that if there is a drug capable of inhibiting 5-lipoxygenase, a parent enzyme of the biosynthesis system of these leukotrienes, then effective prevention and curative effects of various diseases caused by the excessive occurrence of leukotrienes (e.g. allergic diseases, bronchial asthma, hydropic swelling, various inflammatory diseases) can be expected.

On the other hand, in the arachidonate cascase there is the metabolic pathway called 12-lipoxygenase. 12-lipoxygenase is an enzyme that exists mainly in platelets and forms 12-hydroperoxyeicosatetraenoic acid (hereinafter referred to as 12-HPETE) following a reaction with arachidonate, the compound after reduction, 12-hydroxyeicosatetraenoic acid (hereinafter referred to as 12-HETE) is formed.

The physiological significance of the metabolic products for the 12-lipoxygenase pathway is not yet clear compared to that of the 5-lipoxygenase pathway; however, numerous physiological activities of the metabolic products have been recently elucidated with 12-HPETE and 12-HETE as the main metabolic products at the center.

These physiological activities can be exemplified as follows. Namely, with respect to the metabolic products of 12-lipoxygenase, there is a possibility that it may participate in arteriosclerosis by accelerating functional control, such as the aggregation and adhesion of platelets and the migration of vascular smooth muscle cells ("Gendai Iryo", Vol. 21, No. 11, pp. 3109...3113, 1989); one possibility has been suggested in that 12-HPETE may be an initiator for the occurrence of vasospasm after subarachnoid hemorrhage ("Gendai Iryo", Vol. 21, No. 11, pp. 3127...3130, 1989); Furthermore, 12-HETE has been shown to accelerate the adhesion and metastasis of some cancer cells to vascular endothelial cells ("Gendai Iryo", Vol. 22, Special Issue, pp. 56...57, 1990).

Based on the above facts, it is assumed that substances that inhibit 12-lipoxygenase can be used effectively as drugs for the prevention and treatment of various circulatory diseases, such as arteriosclerosis and vascular spasms, but also for the prevention of metastasis of some carcinomas.

Baicalein, a type of natural flavonoid, is known as a substance with the effect of 12-lipoxygenase inhibition (“Biochemical and Biophysical Research Communications”, Vol. 105, No. 3, pp. 1090...1095, 1982).

In addition, the following are known: a hydroxamic acid derivative (Official Gazette "Japanese Patent Kokai Publication" No. 216961/1989, Official Gazette "Japanese Patent Kokai Publication No. 75 2/1990, Official Gazette "Japanese Patent Kokai Publi cation, No. 196767/1990); and caffeic acid derivative (Official Gazette "Japanese Patent Kokai Publication, No. 275552/1989, Official Gazette "Japanese Patent Kokai Publication No. 235852/1990)

On the other hand, a stilbene derivative and an alpha,beta-diarylacrylonitrile derivative are generally synthesized by chemical means or separated and purified from naturally occurring substances; as compounds similar to the compounds of the present invention, the following 1) to 6) are known:

1) alpha-[(3,4-Dihydroxyphenyl)methylene]-4-nitrobenzenelacetonitrile ("Journal of the Society of Dyers and Colourists", Vol. 92, No. 1, p. 14, 1976),

2) alpha-[(3,4-dihydroxy-5-nitrophenyl)methylene)-2-pyrimidine acetonitrile ("Journal of Computer-Aided Molecular Design", Vol. 6, No. 3, p. 253, 1992),

3) alpha-[(3,4-dihydroxyphenyl)methylene]-3,4-dihydroxybenzeneacetonitrile (described in US-P-4,015,017),

4) 2-(p-azidophenyl)-3-(3,4-dihydroxyphenyl)acetonitrile (description in FR-P-1 513 907),

5) alpha'-cyano-3',4'-dihydroxy-4-stilbenecarboxylic acid (description of US-P-2,766,271),

6) alpha'-(diphenylmethylene)-3,4-dihydroxybenzeneacetonitrile (description of DE-A-25 01 443).

However, it has now become known that these compounds collectively possess 12-lipoxygenase inhibition activities.

Furthermore, 12-lipoxygenase is an enzyme related to 5-lipoxygenase, and a substance that can inhibit one of the enzymes also inhibits the other. In fact, a substance is known that, according to publications, has 5-lipoxygenase inhibition activities and also has 12-lipoxygenase inhibition activities, most hydroxamic acid derivatives being examples of this.

Regarding the selectivity of the inhibitory activities, substances that can inhibit 12-lipoxygenase strongly and selectively are preferred for the prevention and cure of diseases, such as above-mentioned circulatory diseases and metastases, which are believed to be caused mainly by metabolic 12-lipoxygenase products, although this depends on the specific object of use.

Disclosure of the invention

Due to this situation, the inventors of the present invention have found that baicalein, a kind of naturally occurring flavonoid, has relatively strong 12-lipoxygenase inhibition activities, has relatively high selectivity, and have produced compounds having strong and highly selective 12-lipoxygenase inhibition activities by using the compound as a lead compound and making the modification of its partial structure, thereby achieving the present invention.

Furthermore, when a compound is used as a drug, for example as an oral drug, one of the crucial preconditions is generally that the compound is effectively absorbed into the body. Crucial factors influencing effective absorption include properties such as hydrophobicity and polarity of the compound.

The degree of hydrophobicity and polarity of the above compound vary greatly, as can be expected from the variety of substituents. Therefore, when the compound is used, for example, as a drug for oral administration, it does not always have the hydrophobicity and polarity suitable for absorption from the intestinal tract.

In connection with the numerous modifications of these compounds, the inventors of the present invention have investigated so-called precursors of drugs which can be effectively absorbed in vivo by imparting to them the hydrophobicity or polarity suitable as drugs and which form primary compounds by being cleaved from the modified part of the compounds rapidly or slowly according to the tasks via the function of the enzyme present in vivo after absorption, as a result of which it was found that a monoacyl substance and a diacyl substance of a catechol hydroxyl group which these compounds usually have are a group of compounds suitable for the above-mentioned task and which has led to the object of the present invention.

An object of the present invention is to provide a compound capable of inhibiting 12-lipoxygenase with a strong and high selectivity.

Another object of the present invention is to provide novel compounds of a stilbene derivative and a stilbene homologue derivative which are useful as so-called "pro-drugs" and which can be effectively absorbed in vivo and can form 12-lipoxygenase inhibiting compounds by cleaving off the modified part in vivo.

Another object of the present invention is to provide drugs with selective 12-lipoxygenase inhibition which can be used as drugs for the prevention and cure of various circulatory diseases such as arteriosclerosis and vasospasms caused by metabolic products of 12-lipoxygenase, as well as for the formation of metastases of some carcinomas, and which, in addition, have low toxicity and few side effects.

A first embodiment of the present invention which solves the above problems is a stilbene derivative or a stilbene homologue derivative represented by the following general formula 9:

(wherein R1 represents a hydrogen atom or a hydroxy group; R2 and R3 are hydrogen atoms or cyano groups (R2 and R3 are different from each other); and Ar is a group represented by the following general formula 10, 11 or 12:

wherein R⁴ represents a hydrogen atom, a lower alkyl group, a lower alkoxy group, a halogen atom, a trifluoromethyl group or a cyano group).

A second embodiment of the present invention which solves the above problems is a pharmaceutical composition containing a compound selected from a group consisting of a sylbene derivative and a stilbene homologue derivative, or a mixture thereof as an effective component and represented by the following formula 13:

(wherein R1 represents a hydrogen atom or a hydroxy group; R2 and R3 are hydrogen atoms or cyano groups (R2 and R3 are different from each other); and Ar is a group represented by the following general formula 14, 15 or 16:

wherein R4 represents a hydrogen atom, a lower alkyl group, a lower alkoxy group, a halogen atom, a trifluoromethyl group, a cyano group or a nitro group).

The compound of the first embodiment of the present invention is stilbene or such a stilbene homologue in which one of the benzene rings of the stilbene is replaced by a heterocyclic ring represented by the following general formula 17:

and relates to a sylbene derivative or stilbene homologue derivative, characterized in that it has at least two hydroxy groups of the catechol type in the A-ring, has no hydroxy group in the B-ring and at each carbon of the Double bond connecting the A ring and the B ring has a cyano group bound.

A method for establishing the connection in conjunction with the first embodiment of the present invention can be exemplified as follows.

It can be a compound represented by the following general formula 18:

(wherein R is a hydrogen atom or a hydroxy group; Ar is a group represented by the general formulas 19, 20 or 21:

wherein R⁴ represents a hydrogen atom, a lower alkyl group, a lower alkoxy group, a halogen atom, a trifluoromethyl group, a cyano group or a nitro group; hereinafter Ar is referred to merely as "Ar") can be prepared synthetically by the processes represented by the following formulas:

(wherein R represents a hydrogen atom or a hydroxy group, R' is a hydrogen atom or OMe).

The compound (III) can be obtained by condensing an intended benzaldehyde derivative (I) and arylacetonitrile (II) under the reaction conditions known as Knoevenagel condensation (process I). Subsequently, the intended compound can be obtained by reacting the aforementioned compound with a demethylated reagent such as boron tribromide, trimethylsilyl iodide or pyridinium chloride (process II).

When the arylacetonitrile (II) to be used in the condensation reaction has a relatively high reactivity, the desired compound can be obtained by subjecting the compound and hydroxybenzaldehyde (IV) to a Knoevenagel condensation reaction according to the process shown in the following reaction scheme (Process III):

Furthermore, the compound represented by the following general formula: Formula 24:

(wherein R is hydrogen or a hydroxy group, can be prepared synthetically by the processes represented by the following formulas: Process IV and V:

wherein R represents a hydrogen atom or a hydroxy group; and R' is a hydrogen atom or OMe).

The compound (VII) can be obtained by reacting an intended phenylacetonitrile derivative (V) and aromatic aldehyde (VI) under the reaction conditions known as Knoevenagel condensation (Process IV). Thereafter, the intended compound can be obtained by demethylating the compound in the same manner as in the aforementioned Process II (Process V).

The compounds of the present invention thus obtained can be purified by known purification methods, such as, for example, recrystallization or chromatography. Although in the compounds of the present Although cis- and trans-geometric isomers exist in the context of the present invention, all of these are otherwise included in the compounds of the present invention.

Since the above-mentioned compounds of the present invention have selective 12-lipoxygenase inhibition, they have effects of inhibiting the formation of 12-lipoxygenase metabolic products such as 12-HPETE and 12-HETE, the pharmaceutical compositions of the present invention containing these compounds as effective components and selectively inhibiting 12-lipoxygenase can be used as pharmaceutical compositions for preventing and curing various circulatory diseases such as arteriosclerosis and vascular spasms, and for effectively preventing metastasis of some carcinomas.

A third embodiment of the present invention is a stilbene derivative and a stilbene homologue derivative represented by the following general formula: Formula 15b:

(wherein R¹ represents a hydrogen atom or OZ; R2 and R3 are hydrogen atoms or cyano groups (R2 and R3 are different from each other); and Ar is a group represented by the following general formula 16b, 17b or 18b:

wherein R4 represents a hydrogen atom, a lower alkyl group, a lower alkoxy group, a halogen atom, a trifluoromethyl group, a cyano group or a nitro group; z is independently a hydrogen atom or a group represented by the general formula 19b (all Z's being other than hydrogen atoms): Formula 19b:

wherein R5 represents a straight-chain or branched-chain alkyl group or alkenyl group having 1 to 20 carbon atoms.

A fourth embodiment of the present invention which solves the above problems is a pharmaceutical composition containing a compound selected from a group consisting of a stilbene derivative and a stilbene homologue derivative or a mixture thereof as an effective component, represented by the following general formula: Formula 20b:

(wherein R¹ represents a hydrogen atom or OZ; R2 and R3 are hydrogen atoms or cyano groups (R2 and R3 are different from each other); and Ar is a group represented by the following general formula 21b, 22b or 23b: Formulas 21b-23b:

wherein R⁴ represents a hydrogen atom, a lower alkyl group, a lower alkoxy group, a halogen atom, a trifluoromethyl group, a cyano group or a nitro group; z is independently a hydrogen atom or a group represented by the general formula 24b (all Z are not hydrogen atoms): Formula 24b:

wherein R⁵ represents a straight-chain or branched-chain alkyl group or alkenyl group having 1 to 20 carbon atoms).

A method for producing the compound (precursor) of the third embodiment of the present invention can be exemplified as follows. Namely, the compound of the present invention can be synthetically produced according to the processes represented by the following reaction schemes: Process A Process B Process C

In the above formulas, R7 represents a hydrogen or a methoxy group; R8 is a hydrogen atom or a hydroxy group, R9 is a hydrogen atom or OZ; Z is a group represented by the general formula 27b:

wherein R10 represents a straight-chain or branched-chain alkyl group or alkenyl group having 1 to 20 carbon atoms; Ar is a group represented by the general formula 28b, 29b or 30b: Formulas 28b...30b:

wherein R¹¹ represents a hydrogen atom, a lower alkyl group, a lower alkoxy group, a halogen atom, a trifluoromethyl group, a cyano group or a nitro group.

The compound (e) or (f) can be prepared by condensing 3,4-dimethoxybenzaldehyde or 3,4,5- trimethoxybenzaldehyde (a) and the desired arylaceitonitrile (b) or 3,4-dimethoxyphenylacetonitrile or 3,4,5-trimethoxyphenylacetonitrile (c) and the desired aromatic aldehyde (d) under the reaction conditions known as Knoevenagel condensation (Process A).

Thereafter, the compound (g) or (h) can be obtained by reacting these compounds with a demethylating agent such as pyridinium chloride (process B). Further, the compound (i) or (j) of the present invention can be obtained by reacting these compounds with an intended acylating agent such as acid chloride and acid anhydride in the presence of a base such as trimethylamine or pyridine. In this case, partially acylated compounds can be obtained by regulating the equivalence relationship of an acylating agent (process C).

If the arylacetonitrile (b) in process A has a relatively high reactivity, the Compounds can also be obtained by the process represented by the following formulas:

In the above formulas, R8 represents a hydrogen atom or a hydroxy group.

Incidentally, the compound (g) can be obtained by directly subjecting 3,4-dihydroxybenzaldehyde or 3,4,5-trihydroxybenzaldehyde (k) instead of 3,4-dimethoxybenzaldehyde or 3,4,5-trimethoxybenzaldehyde and the desired arylacetonitrile (b) to Knoevenagel condensation.

As an acyl group for a modified part of the compounds of the present invention, those having different carbon numbers and different groups including monoacyl, diacyl and triacyl groups can be used to prepare compounds having the desired hydrophobicity and polarity. As for an acyl group, it is not particularly limited in that it is absorbed and cleaved in vivo and its generating free acid can be medically and pharmaceutically accepted; however, when an acyl group is cubically bulky, the decrease in the hydrolysis rate due to an esterase is observed. Thus, there is also the possibility of using the above-mentioned fact to control the cleavage of the compounds of the present invention in vivo by selecting the type of an acyl group as a modified part.

The compounds of the present invention thus obtained can be purified by a known purification method such as recrystallization or chromatography. The compounds of the present invention purified here are remarkably stable and can exist without any change for a long period of time even when they come into contact with an aqueous solution having a pH of 1 to 8. In the compounds of the present invention, cis and trans geometric isomers exist, all of which are otherwise included in the compounds of the present invention.

The above-mentioned compounds of the present invention are absorbed in vivo, acyl groups are cleaved due to the function of the enzymes present in vivo, as a result of which the produced compounds have a strong selective 12-lipoxygenase inhibition and thus an activity for inhibiting the production of 12-lipoxygenase metabolic products such as 12-HPETE and 12-HETE, thus being useful as drugs for the cure and prevention of various circulatory diseases such as arteriosclerosis and vasospasm caused by these metabolic products, and for the effective prevention of metastasis of some carcinomas.

A fifth embodiment of the present invention which solves the above problems is an alpha,beta-diarylacrylonitrile derivative represented by the following general formula 11c: Formula 11c:

(wherein R1 represents a hydrogen atom or a hydroxy group; R2 and R3 are hydrogen atoms or cyano groups (R2 and R3 are different from each other); and Ar is a group represented by the following general formula 12c or 13c: Formula 12c and 13c:

where X represents an oxygen atom or a sulfur atom ).

A sixth embodiment of the present invention which solves the above problems is a pharmaceutical composition containing a compound as an effective component selected from a group consisting of alpha,beta-diarylacrylonitrile derivatives represented by the following general formula 14c, or a mixture thereof: Formula 14c:

(wherein R¹ represents a hydrogen atom or a hydroxy group; R2 and R3 are hydrogen atoms or cyano groups (R2 and R3 are different from each other); and Ar is a group represented by the following general formula 15c or 16c: Formula 15c, 16c:

where X represents an oxygen atom or a sulfur atom ).

A process for producing the compound of the fifth embodiment of the present invention can be exemplified as follows. Namely, the compound of the present invention can be synthesized according to the processes represented by the following formulas: Process A Process B

(wherein R6 represents a hydrogen atom or a methoxy group; R7 represents a hydrogen atom or a hydroxy Group; Ar is a group represented by the following general formula 18c or 19c: Formulas 18c, 19c:

where X represents an oxygen atom or a sulfur atom ).

Namely, in the above formulas, the compounds (e) or (f) can be obtained by condensing 3,4- dimethoxybenzaldehyde or 3,4,5-trimethoxybenzaldehyde (a) and the desired arylacetonitrile (b) or 3,4-dimethoxyphenylacetonitrile or 3,4,5-trimethoxyphenylacetonitrile (c) and the desired aromatic aldehyde (d) under the reaction conditions known as Knoevenagel condensation (process A). Thereafter, the alpha,beta-diarylacrylonitrile derivative compounds (g) or (h) of the present invention can be obtained by reacting these compounds with a dimethylating agent such as pyridinium chloride (process B).

In this case, if arylacetonitrile (b) has a relatively strong reactivity in process A, the compounds can also be obtained by the process represented by the following formulas (process C): Process C

In the above formulas, R7 represents a hydrogen atom or a hydroxy group.

Namely, the compound of the alpha,beta-diarylacrylonitrile derivative (g) of the present invention can be directly obtained by using 3,4-dihydroxybenzaldehyde or 3,4,5-trihydroxybenzaldehyde (i) instead of 3,4-dimethoxybenzaldehyde or 3,4,5-trimethoxybenzaldehyde (a) in the process A and subjecting the desired arylacetonitrile (b) to Knoevenagel condensation.

The thus obtained compounds of the present invention can be purified by a known purification method such as recrystallization or chromatography. In the compounds of the present invention, cis and trans geometric isomers exist, which are otherwise included in the compounds of the present invention.

The above-mentioned compounds of the present invention have the effect of selective 12-lipoxygenase inhibition and thus have an effect of inhibiting the production of 12-lipoxygenase metabolic products such as 12-HPETE and 12-HETE, so that pharmaceutical compositions of the present invention containing these compounds as effective components and which selectively inhibit 12-lipoxygenase can be used as pharmaceutical compositions for the cure and prevention of various circulatory diseases such as arteriosclerosis and vascular spasms caused by these metabolic products, as well as for the effective prevention of metastasis of some carcinomas.

The compounds according to the invention can be used as they are or as medicaments in appropriate dosage forms, such as tablets, capsules, injection solutions, granules, depot dosage forms or the like, by admixing them with a pharmaceutically acceptable known carrier or excipient or the like.

The pharmaceutical compositions containing the compounds of the invention as active components can be administered orally or parenterally by injection, inhalation, application, or the like. The dose of administration may depend on the patients to be treated, the patients' diseases, age, treatment conditions, or the like, so that administration is preferably usually carried out at a dose in the range of about 0.1 mg to 50 mg once or three times a day.

The present invention will now be described in detail using test examples.

Test example 1

The test was conducted to investigate the inhibitory effect of various composite compounds on the activity of 12-lipoxygenase.

1) Preparation of enzyme solution

A male Sprague Dawley rat was anesthetized with ether and blood was collected using a syringe filled to approximately one-tenth volume with 3.8% sodium citrate. The blood was centrifuged at 180 x g for 15 minutes at room temperature to pellet blood-containing platelet plasma. The resulting supernatant was again centrifuged at 1800 x g for 10 minutes at 4°C. The pelleted platelets were washed twice with a washing buffer solution (50 mM Tris-hydrochloric acid buffer containing 154 mM sodium chloride and 2 mM EDTA, pH: 7.4). The platelets were then resuspended in one-twentieth volume of a resuspension buffer (50 mM Tris-hydrochloric acid buffer containing 154 mmol sodium chloride and 5.5 mmol glucose, pH: 7.4). The platelets were then sonicated at 4ºC and incubated for 30 minutes at 100,000 x g at 4ºC to obtain a 12-lipoxygenase enzyme solution as a supernatant.

2) Procedure for 12-lipoxygenase inhibition assay

The enzyme solution described above was diluted with the above-mentioned resuspension buffer to prepare an assay solution so as to include about 2 mU/ml of 12-lipoxygenase. To an aliquot (300 microliters) of the assay solution were added 1 microliter of a 3 mM indomethacin-ethanol solution, 1 microliter of an aqueous 300 mM reduced glutathione solution and 3 microliters of a solution of test compounds having different concentrations in ethanol prepared in the same manner as in Example 1 and Comparative Examples 1 to 9. The resulting mixture was incubated at 37°C for 5 minutes. Then, 3 microliters of a 2.5 mmol arachidonic acid solution in ethanol was added to incubate for a further 5 minutes at 37°C, after which the reaction was stopped by adding 600 ml of methanol. The 12-hydroxyeicosatetraenoic acid recovered in the supernatant after centrifugation at 10,000 x g for 5 minutes was separated by a reversed-phase high-performance liquid chromatography equipped with a C-18 column, and the absorbance of diene at 234 nm was measured to quantify the activity of 12-lipoxygenase. From the experimental data of each sample, the values representing the concentration to produce 50% inhibition of 12-lipoxygenase activity (hereinafter referred to as IC50) were determined.

3) Test results

The test results are shown in Table 1 and Table 2. In Table 1, the 12-lipoxygenase inhibition ratios of the compounds of Comparative Examples 1 to 4, whose IC50 values could not be obtained, and that of the compound of Example 1 according to the present invention for comparison purposes are shown; while in Table 2 shows the compounds of Comparative Examples 5 to 9, whose IC50 values could be obtained in contrast to the compound (Example 1) of the present invention.

As is clear from Table 1 and Table 2, it was found that although the 12-lipoxygenase inhibitory activity of the compound of Example 1 according to the present invention had an inhibitory rate of 97% at a concentration of 10-5 mol and an inhibitory rate of 73% at a concentration of 10-8 mol, the 12-lipoxygenase inhibitory activity of the compounds of Comparative Examples 1 to 4 had an inhibitory rate of only less than 10% at a concentration of 10-5 mol, although they are similar in chemical structures to the compound of the present invention.

The above results suggest that it is important that the compound of the invention has at least two catichol-type hydroxy groups in ring A, which is represented by the following general formula considering that it exhibits a strong 12-lipoxygenase inhibitory activity: Formula 26:

In addition, it was found that the 12-lipoxygenase IC50 inhibitory activity of the compound having two cyano groups in a double bond linking ring A and ring B (Comparative Example 5), the compound having an ethoxycarbonyl group instead of a cyano group (Comparative Example 6) and the compounds having a hydroxy group or a carboxy group in the B ring (Comparative Examples 7 to 9) was higher than that of of the compound of the invention (Example 1) is remarkably poor. Otherwise, as a result of carrying out a test with the other compounds of the present invention, almost the same results were obtained. Table 1 Results of the test of the inhibition levels Compound 12-lipoxygenase inhibition levels (%)

Note: 1) The numerical values in line 3 show the molar concentrations of the compounds.

2) - indicates that no measurement has been carried out .

Table 2 Results of the inhibitory effect test Compound 12-lipoxygenase IC₅₀ values

Example 1 4.2 x 10&supmin;&sup5;

Compare Example 5 1.2 · 10⊃min;⊃6;

Compare Example 6 1.6 · 10⊃min;⊃7;

Compare Example 7 2.3 · 10⊃min;⊃7;

Compare Example 8 3.0 · 10⊃min;⊃7;

Compare Example 9 5.0 · 10⊃min;⊃6;

Test example 2

The test was carried out to investigate the selectivity towards 12-lipoxygenase of the compounds of the first embodiment of the present invention.

1) Preparation of the enzyme solution i) Preparation of 5-lipoxygenase enzyme solution

Rat basophil leukemia cells (RBL-1, ATCC CRL1378) were grown in Dulbecco's modified Eagle's medium containing 10% newborn bovine serum.

The grown cells were collected and washed twice with 50 ml of Tris-buffered saline (containing 154 mmol sodium chloride, pH 7.4; hereafter referred to as TBS). The cell suspension (4 x 107 cells/ml) was then sonicated and centrifuged at 10,000 x g for 10 minutes.

The resulting supernatant was used to prepare an assay solution.

ii) Preparation of 12-lipoxygenase enzyme solution

The enzyme solution was prepared in the same manner as in Test Example 1.

2) Method of measuring enzyme activity i) Procedure for 5-lipoxygenase inhibition assay

To an aliquot (15 microliters) of the enzyme solution (40 mU/ml equivalent), 185 microliters of TBS, 50 microliters of 2 mmol adenosine triphosphate in TBS, 50 microliters of 12 mmol calcium chloride in TBS, 1 microliter of 3 mmol indomethacin in ethanol solution, 1 microliter of an aqueous reduced 300 mol glutathione solution and 3 microliters of test compounds in ethanol solution of different concentrations were added. The resulting mixture was incubated for 5 minutes at 37ºC. Thereafter, 3 microliters of a 2.5 mmol arachidonic acid solution in ethanol were added to incubate the mixture for 2 minutes at 37ºC. The reaction was stopped by adding 600 microliters of methanol, after which it was centrifuged at 10,000 x g for 5 minutes and the 5-hydroxyeicosatetraenoic acid obtained in the supernatant was separated by reversed-phase high performance liquid chromatography equipped with a C-18 column, after which the absorbance of diene was measured at 234 nm to quantify the activity of the enzyme.

Using the compounds of the invention prepared in the same manner as in Examples 1 to 6, Example 9, Example 11, Examples 13 to 17 and Examples 20 to 27 as test samples and known baicalein (purchased from Wako Junyaku Kogyo) represented by the following formula as a control, the tests for dose-dependent inhibition were carried out to determine the IC50 values: Formula 27:

xii) Procedure for 12-lipoxygenase inhibition assay

The inhibition assay was carried out according to the same method as described in Test Example 1, except that the same samples were used to measure 5-lipoxygenase activity as were used to determine IC50 values.

3) Test results

The results of the test are shown in Table 3. As can be seen from Table 3, the compounds of the present invention revealed a remarkably strong inhibitory effect on 12-lipoxygenase, and the IC50 values were on the order of 10-9 mol, which was equivalent to or better than the known baicalein. On the other hand, the compounds of the present invention also had an inhibitory effect on 5-lipoxygenase, and the IC50 values were 10 to 30 times or more of those of 12-lipoxygenase. This showed that the compounds of the present invention had a selective inhibitory effect on 12-lipoxygenase.

Among the compounds of the present invention, those having a relatively large substituent in the B ring tend to have a stronger 12-lipoxygenase inhibitory activity, wherein when the B ring is a benzene ring, compounds having a substituent in the p-position are preferred, while when the B ring is a thiophene ring or a furan ring, those having a substituent Ente is preferred which is in the m-position to the substitution site of a double bond linking the A-ring.

A particularly remarkable 12-lipoxygenase inhibitory effect was shown by substituents as a chlorine atom or a bromine atom in the case of a halogen atom, as a methyl group or an ethyl group in the case of an alkyl group, and as a methoxy group in the case of an alkoxy group.

Furthermore, the results obtained when carrying out the test on other compounds according to the invention were largely the same. Table 3 Test results with regard to the selectivity of an inhibitory effect

Test example 3

The test was carried out to investigate the NMR spectra and the IR absorption spectra of the compound of the first Embodiment according to the present invention and the comparison compounds.

1) Preparation of samples

The compounds of the invention were prepared in the same manner as in Examples 1 to 17 and Examples 20 to 29, and the compounds for comparison were prepared in the same manner as in Comparative Examples 1 to 9.

2) Test method

The NMR spectra (1H-NMR (500 MHz)) were measured in a CDCl3 solvent to which a few drops of CD3OD were added; the IR absorption spectra were measured by a KBr tablet method.

3) Test results

The results of the test are shown in Tables 4 to 11. Table 4 Table 5 Table 6 Table 7 Table 8 Table 9 Table 10 Table 11

(Comparison example 1)

The yellow crystalline compound, 551 mg (yield: 43.1%) represented by the following formula was obtained by the same procedure as in Example 1 except that 611 mg (5.0 mmol) of 3-hydroxybenzaldehyde (purchased from Tokyo Kasei) was used instead of 3,4-dihydroxybenzaldehyde: Formula 28:

The melting point of the obtained compound was 114.5ºC ... 116.5ºC.

(Comparison example 2)

The yellow crystalline compound, 975 mg (yield: 76.2%) represented by the following formula was obtained by the same procedure as in Example 1 except that 611 mg (5.0 mmol) of 4-hydroxybenzaldehyde (purchased from Tokyo Kasei) was used instead of 3,4-dihydroxybenzaldehyde: Formula 29:

The melting point of the obtained compound was 200ºC ... 202ºC.

(Comparison example 3)

The pale yellow crystalline compound, 488 mg (yield: 97.6%) represented by the following formula was obtained by the same procedure as in Example 1 except that 761 mg (5.0 mmol) of 3-hydroxy-4-methoxybenzaldehyde (purchased from Jansen) was used instead of 3,4-dihydroxybenzaldehyde: Formula 30:

The melting point of the obtained compound was 139ºC ... 140ºC.

(Comparison example 4)

The yellow crystalline compound, 481 mg (yield: 68.7%) represented by the following formula was obtained by the same procedure as in Example 1 except that 761 mg (5.0 mmol) of f4-hydroxy-3-methoxybenzaldehyde (purchased from Tokyo Kasei) was used instead of 3,4-dihydroxybenzaldehyde: Formula 31:

The melting point of the obtained compound was 122ºC ... 1213ºC.

(Comparison example 5)

To a suspension of benzyltriphenylphosphonium bromide (4.33 g, 10 mmol, purchased from Lancaster) in 12 ml of tetrahydrofuran (purchased from Aldrich) was added a Solution of lithium bistrimethylsilylamide (1.0 mol solution in tetrahydrofuran, 10 mL, purchased from Aldrich) was added at room temperature and the mixture was stirred for an additional 30 minutes. To the mixture was added a solution of 3,4-dimethoxybenzaldehyde (1.5 mg, 9 mmol, purchased from Tokyo Kasei) in 6 mL of tetrahydrofuran and the resulting mixture was stirred for an additional 2.5 hours. The reaction was quenched by addition of methanol. The crude product obtained after usual treatment was subjected to chromatography on silica gel to give a mixture of trans and cis isomers of 3,4-dimethoxystilbene. The mixture was recrystallized from hexane-ethyl acetate (30:1) to give 680 mg (yield: 31.0%) of the trans isomer.

A mixture of 222 mg (0.93 mmol) of the above-mentioned trans-3,4-dimethoxystilbene and 1.5 g (13 mmol) of pyridinium chloride (purchased from Wako Junyaku Kogyo) was melted and stirred at 200 °C in an argon gas atmosphere for 45 minutes, air-cooled, treated with 50 ml of 2N hydrochloric acid, extracted with ethyl acetate, washed with saturated brine and dried over anhydrous magnesium sulfate. The solvent was evaporated under reduced pressure and the residue was subjected to thin layer chromatography (ethyl acetate/hexane = 1:1) to obtain 100 mg (yield: 50.0%) of a white crystalline compound represented by the following formula: Formula 32:

The melting point of the obtained compound was 170... 171ºC.

(Comparison example 6)

To a mixture of ethyl phenylacetate (821 mg, 5.0 mmol, purchased from Tokyo Kasei) and 3,4-dihydroxybenzaldehyde (691 mg, 5.0 mmol, purchased from Tokyo Kasei) in 10 mL of ethanol was added 0.54 mL of piperidine (purchased from Wako Junyaku Kogyo) and refluxed for 24 hours. After cooling, the mixture was poured into 100 mL of 1N hydrochloric acid. The precipitate was collected and washed with water and then dissolved in ethyl acetate. The solution was washed twice with 20% aqueous sodium hydrogen sulfite and then with brine. The organic layer was dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure. The residue was purified by column chromatography on silica gel. Elution with hexane/ethyl acetate (10 : 1 ... 10 : 4) gave 499 mg (yield: 35.1%) of the yellow crystalline product represented by the following formula: Formula 33:

The melting point (hereinafter referred to as mp) was 153ºC ... 154.5ºC.

(Comparison example 7)

Following the same procedure as in Example 2, alpha-cyano-3',4,4'-trimethoxystilbene (white crystalline), 2.61 g (yield: 88.0%) was obtained except that 1.47 g (10 mmol) of 4-methoxyphenylacetonitrile (purchased from Aldrich) was used instead of 3,4-dimethoxyphenylacetonitrile and that 1.66 g (10 mmol) of 3,4-dimethoxybenzaldehyde (purchased from Tokyo Kasei) was used instead of 4-chlorobenzaldehyde.

Yellow crystals, 588 mg (yield: 93.0%), represented by the following formula were obtained by the same procedure as in Example 1 except that 0.74 g (2.5 mmol) of the above-mentioned alpha-cyano-3',4,4'-trimethoxystilbene and 3.47 g (30 mmol) of pyridinium chloride (purchased from Wako Junyaku Kogyo) were used: Formula 34:

The melting point of the obtained compound was 229ºC. ...231ºC.

(Comparison example 8)

Alpha-cyano-3,3',4,4'-tetramethoxystilbene (yellow crystalline), 2.99 g (yield: 92.0%), was obtained by the same procedure as in Example 2 except that 1.66 g (10 mmol) of 3,4-dimethoxybenzaldehyde (purchased from Tokyo Kasei) was used instead of 4-chlorobenzaldehyde.

Yellow crystals, 138 mg (yield: 20.5%) represented by the following formula were obtained by the same procedure as in Example 2 except that 0.81 g (2.5 mmol) of the above-mentioned alpha-cyano-3',3,4,4'-tetramethoxystilbene and 3.47 g (30 mmol) of pyridinium chloride (purchased from Wako Junyaku Kokyo) were used: Formula 35:

The melting point of the obtained compound was 238 ... 239 °C.

(Comparison example 9)

A mixture of 3,4-dimethoxyphenylacetonitrile (475 mg, 2.68 mmol, purchased from Tokyo Kasei) and 4-formylbenzoic acid (402 mg, 2.68 mmol, purchased from Tokyo Kasei) was heated to dissolve it in 5 mL of anhydrous ethanol. To the mixture was added a solution of sodium ethoxide (2.68 mol, denatured alcohol, 2 mL, purchased from Aldrich) and the mixture was allowed to stand for one hour. To the mixture was added 30 mL of water and the mixture was stirred. The insoluble material was collected and washed with water and then treated with 30 mL of 2N hydrochloric acid. The precipitate was collected and washed thoroughly with water to give 605 mg (yield: 70.3%) of alpha'-cyano-3',4'-dimethoxy-4-stilbenecarboxylic acid as a yellow crystalline compound.

A mixture of the above-mentioned alpha'-cyano-3',4'-dimethoxy-4-stilbenecarboxylic acid (309 mg, 1.0 mmol) and pyridinium chloride (1.73 g, 15 mmol, purchased from Wako Junyaku Kogyo) was melted and stirred in an atmosphere of argon gas at 200 °C for one hour. After cooling, 2N hydrochloric acid was added to the resulting mass and the mass was crushed and stirred. The precipitate was collected and washed with water and then taken up in ethanol. The solvent was removed under reduced pressure and the residue was recrystallized from ethanol/hexane to give 198 mg (yield: 70.4%) of the yellow crystalline product represented by the following formula, m.p. 285ºC ... 287ºC. Formula 36:

Test example 4

The test was carried out to examine the 12-lipoxygenase inhibitory activity of the compound produced from the compound of the third embodiment of the present invention and absorbed in vivo and then deacylated.

1) Preparation of enzyme solution

This was produced using the same procedure as in Test Example 1.

2) Method of measuring enzyme activity

The measurement was carried out according to the same procedure as in Test Example 1.

3) Test results

The test results are shown in Table 12. In Table 12, the 12-lipoxygenase IC50 values of the compounds produced from the compounds of Examples 30 to 39, 43, 45 and 48 of the present invention, which were absorbed in vivo and then deacylated, were compared in the baicalein (purchased from Wako Junyaku Kogyo) as a known compound for comparison.

As is clear from Table 12, the 12-lipoxygenase inhibitory activity of the compound of each example of the present invention is far superior to that of baicalein. Incidentally, as a result, when a test of the other compounds of the present invention was carried out, almost the same results were obtained. Table 12

Note: * ... IC₅₀₀ values

Test example 5

The test was carried out to examine whether the deacylated compounds are formed due to esterases present in the enzyme solution when the compounds in the third embodiment of the present invention are treated with the same enzyme solution as in Test Example 4.

1) Preparation of enzyme solution

The enzyme solution was prepared following the same procedure as described in Test Example 4.

2) Test method

1 ml of the above enzyme solution was diluted with 11 ml of a resuspension buffer. An aliquot (300 To 3 microliters of a 1.0 x 10-3 molar solution of a test compound in dimethyl sulfoxide was added (100 microliters) of this enzyme solution. The mixture was incubated at 37°C for the indicated time, after which the reaction was stopped by adding 1 ml of acetonitrile. 20 ml of the above mixture was subjected to reversed-phase high performance liquid chromatography equipped with a C-18 column to quantify the amount of deacylated products. As a control, an assay was carried out using 300 microliters of the above resuspension buffer solution in place of the enzyme solution.

The conditions of the reverse phase high performance liquid chromatography were:

Column: Superspher RP-18(e) (purchased from Merck), Diameter: 4.0 mm, Length: 125 mm

Flow rate: 1.0 ml/min

Eluent: Acetonitrile containing 50 mmol NaClO4 and

50 mMol orthophosphate/water (55 : 45)

Column temperature: 30ºC

Calibration: Extinction of 280 nm

3) Test results

The test results of the compounds of Example 30 (Formula 34b) and Example 31 (Formula 35b) of the compounds of the present invention are shown. Since both of the compounds of Example 30 (Formula 34b) and Example 31 (Formula 35b) were considered to be converted into the compound of Reference Example 1 (Formula 32b) by deacylation, the retention time of the compound of Reference Example 1 was checked exactly under the conditions used by the reversed-phase high performance liquid chromatography as mentioned above (retention time: 4.06 minutes) and a calibration curve of the compound was prepared using the authentic sample of Reference Example 1.

As a case of using the resuspension buffer instead of the enzyme solution, neither of the two Compounds of Example 30 (Formula 34b) and Example 31 (Formula 35b) produce a deacylated compound (Formula 32b).

When the similar test was carried out using the above enzyme solution, the two compounds were deacylated at increased times, and the deacylated compound was detected in the reaction solution as shown in Table 13.

These results clearly demonstrate that modified moieties (referred to herein as "unsubstituted groups") in the compounds of the invention were cleaved relatively rapidly to form a deacylated compound when contacted with an enzyme solution (possibly containing some type of esterase in addition to 12-lipoxygenase) prepared from rat platelets.

On the other hand, the compounds produced by deacylation showed a remarkably strong inhibitory activity against 12-lipoxygenase, as shown in Test Example 4. The compounds described in the present invention were considered as precursors for the production of compounds having inhibitory activity against 12-lipoxygenase and can therefore be used as so-called "por drugs".

When testing other compounds of the present invention, predominantly the same results were obtained. Table 13

Note: The numerical values show a conversion in percent from Formula 34b or Formula 35b to Formula 32b.

Test example 6

The test was designed to investigate the 12-lipoxygenase and 5-lipoxygenase inhibitory effects of the compounds of fifth embodiment of the present invention.

1. Preparation of samples 1) Samples

The samples were prepared by the same procedure as in Examples 54 to 58 to be described later. Besides, as a control, baicalein (purchased from Wako Junyaku Kogyo) was used as a known compound represented by the following formula 21c: Formula 21c:

2) Preparation of 12-lipoxygenase enzyme solution

This was produced using the same procedure as in Test Example 1.

2') Preparation of 5-lipoxygenase enzyme solution

This was produced using the same procedure as in Test Example 2.

2. Test method 1) Method of measuring 12-lipoxygenase enzyme activity

The measurement was carried out according to the same procedure as in Test Example 1.

2) Method of measuring 5-lipoxygenase enzyme activity

The measurement was carried out according to the same procedure as in Test Example 2.

3) Measurement of enzyme activities

The enzyme activities of the compounds of the invention, which were prepared according to the same procedure to be described as in Examples 54 to 58, as well as baicalein as sample substances with different concentrations, were determined according to the method described above. method, whereby the molar concentrations showing 50% 12-lipoxygenase and 5-lipoxygenase inhibition rates (hereinafter referred to as IC50 values) were obtained from the measured values of the crude substances.

3. Test results

The test results are shown in Table 23. As is clear from Table 23, the compounds of the present invention were found to have a remarkably strong inhibitory effect on 12-lipoxygenase, with IC50 values of the order of 10-9 mol being equal to or better than those of the known baicalein. On the other hand, the compounds of the present invention also have an inhibitory effect on 5-lipoxygenase, with IC50 values of the order of 3 to 30 times or more better than those on 12-lipoxygenase. Thus, it was demonstrated that the compounds of the present invention selectively have an inhibitory effect on 12-lipoxygenase. Table 23

Best mode for carrying out the invention

Hereinafter, the present invention will be described in detail by means of examples and reference examples; the present invention is in no way limited to the following examples.

example 1

To a mixture of 4-chlorophenylacetonitrile (758 mg, 5.0 mmol, purchased from Tokyo Kasei) and 3,4-dihydroxybenzaldehyde (691 mg, 5.0 mmol, purchased from Tokyo Kasei) in 10 mL of ethanol was added 0.54 mmol of piperidine (purchased from Wako Junyaku Kogyo) and refluxed for 6 hours. After cooling, the mixture was poured into 100 mL of 1 N hydrochloric acid and stirred. The precipitate was collected and washed with water and then dissolved in ethyl acetate. The solution was washed twice with 20% aqueous sodium hydrogen sulfite solution and then with brine. The organic layer was dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure. The residue was purified by column chromatography on silica gel. Elution with hexane/ethyl acetate (5 : 1 ... 5 : 2) gave 919 mg (yield: 67.6%) of the yellow crystalline product represented by the following formula with mp 160ºC ... 162ºC: Formula 37:

Example 2

A mixture of 3,4-dimethoxyphenylacetonitrile (1.77 g, 10.0 mmol, purchased from Tokyo Kasei) and 4-chlorobenzaldehyde (1.41 g, 10.0 mmol, purchased from Tokyo Kasei) in 5 mL of ethanol was heated until dissolved. Two drops of 20% aqueous sodium hydroxide solution were added and the reaction mixture was stirred overnight. The resulting reaction mass was ground in ethanol. The precipitate was collected and washed with ethanol and subsequently with hexane and dried to give 2.39 g (yield: 79.7%) of greenish-yellow, to give crystalline alpha-cyano-3,4-dimethoxy-4'-chlorostilbene.

A mixture of the above-mentioned alpha-cyano-3,4-dimethoxy-4'-chlorostilbene (0.75 g, 2.5 mmol) and pyridinium chloride (2.31 g, 20.0 mmol, purchased from Wako Junyaku Kogyo) was melted to mix on an oil bath (210 °C) under an argon atmosphere. Stirring was continued for an additional hour. After cooling, 2N hydrochloric acid and ethyl acetate were added to the resulting reaction mass. The organic layer was separated and the water layer was extracted with ethyl acetate. The combined organic layer was washed with brine and then dried over anhydrous sodium sulfate. The solvent was evaporated under reduced pressure. The residue was dissolved in a small amount of ethyl acetate. Then, hexane was added to the solution until the mixture became slightly turbid and the mixture was stirred. The precipitate was collected and dried to give 455 mg (yield: 67.0%) of the yellow crystalline product represented by the following formula with mp 171ºC ... 172ºC: Formula 38:

Example 3

The light yellow crystalline compound represented by the following formula, 405 mg (yield: 31.7%), was obtained by the same procedure as in Example 1 except that 676 mg (5.0 mmol) of 4-fluorophenylacetonitrile (purchased from Tokyo Kasei) was used instead of 4-chlorophenylacetonitrile: Formula 39:

The melting point of the obtained compound was 184ºC ... 186ºC.

Example 4

The light yellow crystalline compound represented by the following formula, 1.14 g (yield: 72.2%) was obtained by the same procedure as in Example 1 except that 980 mg (5.0 mmol) of 4-bromophenylacetonitrile (purchased from Tokyo Kasei) was used instead of 4-chlorophenylacetonitrile: Formula 40:

The melting point of the obtained compound was 172ºC ... 175ºC.

Example 5

The yellow crystalline compound represented by the following formula, 1.39 g (yield: 76.4%) was obtained by the same procedure as in Example 1 except that 1.22 g (5.0 mmol) of 4-iodophenylacetonitrile, which was synthesized by the following method, was used instead of 4-chlorophenylacetonitrile: Formula 41:

The melting point of the obtained invention was 197... 198°C.

(Synthetic preparation of 4-iodophenylacetonitrile)

A mixture of p-iodotoluene (4.36 g, 20 mmol, purchased from Tokyo Kasei) and N-bromosuccinimide (3.92 g, 22 mmol, purchased from Tokyo Kasei) in 60 mL of carbon tetrachloride (purchased from Wako Junyaku Kogyo) was refluxed for 4 hours under irradiation of an incandescent lamp to give 2.67 g (yield: 45.0%) of 4-iodobenzyl bromide as a white crystalline substance.

To a hot solution (50 °C) of sodium cyanide (0.49 g, 10.0 mmol, purchased from Kokusan Kagaku) in 10 mL of dimethyl sulfoxide (purchased from Aldrich) was added the above-mentioned 4-iodobenzyl bromide (1.48 g, 5.0 mmol) and the mixture was stirred for 3 hours at ambient temperature. To the resulting reaction mass was added water and extracted with hexane. The solvent was removed to give 0.84 g (yield: 68.9%) of 4-iodophenylacetonitrile as a white crystalline substance.

Example 6

The yellow crystalline compound represented by the following formula, 5.15 mg (yield: 41.0%) was obtained by the same procedure as in Example 1 except that 656 mg (5.0 mmol) of p-toluylacetonitrile (purchased from Tokyo Kasei) was used instead of 4-chlorophenylacetonitrile: Formula 42:

The melting point of the obtained compound was 164ºC ... 165ºC.

Example 7

The light yellow crystalline compound represented by the following formula, 360 mg (yield: 28.7%) was obtained by the same procedure as in Example 1 except that 656 mg (5.0 mmol) of m-toluylacetonitrile (purchased from Aldrich) was used instead of 4-chlorophenylacetonitrile: Formula 43:

The melting point of the obtained compound was 144ºC ... 145ºC.

Example 8

The light yellow crystalline compound represented by the following formula, 214 mg (yield: 16.0%), was obtained by the same procedure as in Example 1 except that 736 mg (5.0 mmol) of 4-methoxyphenylacetonitrile (purchased from Aldrich) was used instead of 4-chlorophenylacetonitrile: Formula 44:

The melting point of the obtained compound was 197ºC ... 198ºC.

Example 9

The light yellow crystalline compound represented by the following formula, 890 mg (yield: 75.0%), was obtained by the same procedure as in Example 1 except that 586 mg (5.0 mmol) of phenylacetonitrile (purchased from Tokyo Kasei) was used instead of 4-chlorophenylacetonitrile: Formula 45:

The melting point of the obtained compound was 148ºC ... 150ºC.

Example 10

The yellow crystalline compound represented by the following formula, 647 mg (yield: 87.4%) was obtained by the same procedure as in Example 1 except that 926 mg (5.0 mmol) of 4-trifluoromethylphenylacetonitrile (purchased from Aldrich) was used instead of 4-chlorophenylacetonitrile: Formula 46:

The melting point of the obtained compound was 168ºC ... 169ºC.

Example 11

The light yellow crystalline compound represented by the following formula, 281 mg (yield: 21.2%), was obtained by the same procedure as in Example 1 except that 726 mg (5.0 mmol) of 4-ethylphenylacetonitrile synthesized by the following method was used instead of 4-chlorophenylacetonitrile: Formula 47:

The melting point of the obtained compound was 137ºC ... 138ºC.

(Synthetic preparation of 4-ethylphenylacetonitrile)

A mixture of 4-ethylbenzyl alcohol (2.72 g, 20 mmol, purchased from Aldrich) and 50 mL of 47% hydrobromic acid (purchased from Wako Junyaku Kogyo) was stirred vigorously for 30 minutes at room temperature. The reaction mixture was extracted with hexane to give 3.98 g of 4-ethylbenzyl bromide as a colorless oil. Then, a hot solution (50 °C) of sodium cyanide (1.96 g, To 20 ml of dimethyl sulfoxide (40 mmol) was added the above-mentioned 3.98 g of 4-ethylbenzyl bromide and the mixture was stirred for 3 hours at ambient temperature. To the resulting reaction mass was added water and extracted with hexane. The solvent was removed to give 2.70 g (yield: 93.1%) of 4-ethylphenylacetonitrile as a pale yellow oil.

Example 12

The yellow crystalline compound represented by the following formula, 281 mg (yield: 21.2%), was obtained by the same procedure as in Example 1 except that 616 mg (5.0 mmol) of thiophen-2-yl-acetonitrile (purchased from Tokyo Kasei) was used instead of 4-chlorophenylacetonitrile: Formula 48:

The melting point of the obtained compound was 180.5ºC ... 181.5ºC.

Example 13

The yellow crystalline compound represented by the following formula, 1.05 g (yield: 65.0%), was obtained by the same procedure as in Example 1 except that 1.01 g (5.0 mmol) of 4-bromothiophen-2-yl-acetonitrile, which was synthesized by the following method, was used instead of 4-chlorophenylacetonitrile: Formula 49:

The melting point of the obtained compound was 222ºC ... 224ºC.

(Synthetic preparation of 4-bromothiophen-2-yl- acetonitrile)

To a solution of 4-bromothiophene-2-carboxaldehyde (9.55 g, 50 mmol, purchased from Aldrich) in 100 mL of ethanol was added sodium borohydride (3.78 g, 100 mmol, purchased from Yoneyama Yakuhin) gradually with cooling in an ice bath. After the addition was completed, the mixture was stirred for an additional 1.5 hours at room temperature. The reaction mixture was acidified with hydrochloric acid and then concentrated to dryness under reduced pressure. Water was added to the residue and extracted with ether to give 9.29 g (yield: 96%) of 4-bromothiophene-2-yl-methanol as an oil.

A mixture of the above-mentioned 4-bromothiophen-2-yl-methanol (4.83 g, 25 mmol) and 63 ml of 47% hydrobromic acid was vigorously stirred for 30 minutes at room temperature. The reaction mixture was extracted with pentane to give 5.31 g (yield: 82.8%) of 2-bromomethyl-4-bromothiophene as a pale yellow oil. The 5.12 g of 2-bromomethyl-4-bromothiophene thus obtained was reacted with sodium cyanide according to the same procedure as in the synthesis of 4-iodophenylacetonitrile of Example 5 to give 1.50 g (yield: 37.0%) of 4-bromothiophen-2-yl-acetonitrile as a white crystalline substance to give the by-product alpha,alpha-bis((4-bromothiophen-2-yl)methyl]-4- bromothiopheneacetonitrile was removed by recrystallization.

Example 14

The yellow crystalline compound represented by the following formula, 216 mg (yield: 13.4%), was obtained by the same procedure as in Example 1 except that 1.01 g (5.0 mmol) of 5-bromothiophen-2-yl-acetonitrile, which was synthesized by the following method, was used instead of 4-chlorophenylacetonitrile: Formula 50:

The melting point of the obtained compound was 185ºC ... 186ºC.

(Synthetic preparation of bromothiophen-2-yl-acetonitrile)

This compound was synthesized using the same method as the synthesis of 4-bromothiophene-2-yl-acetonitrile of Example 13, except that 5-bromothiophene-2-carboxaldehyde (purchased from Aldrich) was used and purification was carried out by high performance liquid chromatography.

Example 15

The light yellow crystalline compound represented by the following formula, 344 mg (yield: 23.9%), was obtained by the same procedure as in Example 1 except that 771 mg (5.0 mmol) of 3,4,5-trihydroxybenzaldehyde (purchased from Aldrich) was used instead of 3,4-dihydroxybenzaldehyde: Formula 51:

The melting point of the obtained compound was 240ºC ... 143ºC.

Example 16

The yellow crystalline compound represented by the following formula, 558 mg (yield: 85.1%), was obtained by the same procedure as in Example 1 except that 355 mg (2.5 mmol) of cyanophenylacetonitrile synthesized by the following method was used instead of 4-chlorophenylacetonitrile, 345 mg (2.5 mmol) of 3,4-dihydroxybenzaldehyde (purchased from Aldrich), 5 ml of ethanol and 0.27 ml of piperidine (purchased from Wako Junyaku Kogyo) and that the reaction product was stirred at room temperature for 6 hours: Formula 52:

The melting point of the obtained compound was 263ºC ... 265ºC.

(Synthetic preparation of cyanophenylacetonitrile)

There was obtained 2.0 g (yield: 51.0%) of 4-bromomethylbenzonitrile (white crystalline substance) by the same method as that of the synthesis of 4-iodophenylacetonitrile in Example 5 except that 2.34 g (20 mmol) of p-toluenonitrile (purchased from Tokyo Kasei) was used. Thereafter, 438 mg (yield: 34.2%) of light yellow crystals of 4-cyanophenylacetonitrile was obtained by the same method as that of the synthesis of 4-iodophenylacetonitrile in Example 5 except that 1.76 g (9.0 mmol) of the above-mentioned 4-bromomethylbenzonitrile was used and purification was carried out by column chromatography on silica gel and recrystallization.

Example 17

The orange crystalline compound represented by the following formula, 1.18 g (yield: 82.3%), was obtained by the same procedure as in Example 1 except that 811 mg (5.0 mmol) of 4-nitrophenylacetonitrile (purchased from Aldrich) was used instead of 4-chlorophenylacetonitrile and that the reaction product was stirred at room temperature and purification was carried out by recrystallization from ethanol/water: Formula 53:

The melting point of the obtained compound was 260ºC ... 262ºC.

Example 18

A mixture of the following composition was prepared per tablet and processed using a tablet press in the usual manner for preparing a medical composition of the present invention:

Compound obtained in Example 1: 30.0 (mg)

Lactose (purchased from Iwaki Saiyaku) 40.0

Grain starch (purchased from Yoshida Seiyaku) 15.0

Magnesium stearate (purchased from Taihei Kagaku) 0.4

Carboxymethylcellulose calcium (purchased from Nichirin Kagaku Kogyo) 20.0

Example 19

A mixture of the following composition was prepared per capsule and placed in a gelatin capsule in the usual manner for the manufacture of a pharmaceutical composition of the present invention:

Compound obtained in Example 1: 30.0 (mg)

Lactose (purchased from Iwaki Saiyaku) 40.0

Fine powdered cellulose (purchased from Nippon Soda) 30.0

Magnesium stearate (purchased from Taihei Kagaku) 3.0

Example 20

Alpha-cyano-3,4-dimethoxystilbene (yellow crystalline substance), 2.62 g (yield: 98.9%), was obtained by the same procedure as in Example 2 except that 1.06 g (10.0 mmol) of benzaldehyde (purchased from Wako Junyaku Kogyo) was used instead of 4-chlorobenzaldehyde.

The yellow crystalline compound represented by the following formula, 253 mg (yield: 53.4%), was obtained by the same procedure as in Example 2 except that 531 mg (2.0 mmol) of the above alpha- Cyano-3,4-dimethoxystilbene was used instead of alpha-cyano-3,4-dimethyl-4'-chlorostilbene: Formula 54:

The melting point of the obtained compound was 172.5ºC ... 173.0ºC.

Example 21

Alpha-cyano-3,4-dimethoxyfluorostilbene (white crystalline substance), 2.33 g (yield: 82.2%), was obtained by the same procedure as in Example 2 except that 1.24 g (10.0 mmol) of 4-fluorobenzaldehyde (purchased from Tokyo Kasei) was used instead of 4-chlorobenzaldehyde.

The white crystalline compound represented by the following formula, 432 mg (yield: 84.6%), was obtained by the same procedure as in Example 2 except that 567 mg (2.0 mmol) of the above-mentioned alpha-cyano-3,4-dimethoxy-4'-fluorostilbene was used instead of alpha-cyano-3,4-dimethoxy-4'-chlorostilbene: Formula 55:

The melting point of the obtained compound was 175.5ºC ... 176.0ºC.

Example 22

Alpha-cyano-3,4-dimethoxy-3'-chlorostilbene (yellow crystalline substance), 2.35 g (yield: 78.3%), was obtained by the same procedure as in Example 2 except that 1.41 g (10.0 mmol) of 3-chlorobenzaldehyde (purchased from Tokyo Kasei) was used instead of 4-chlorobenzaldehyde.

The white crystalline compound represented by the following formula, 436 mg (yield: 80.2%), was obtained by the same procedure as in Example 2 except that 600 mg (2.0 mmol) of the above-mentioned alpha-cyano-3,4-dimethoxy-3'-chlorostilbene was used instead of alpha-cyano-3,4-dimethoxy-4'-chlorostilbene: Formula 56:

The melting point of the obtained compound was 189ºC ... 190ºC.

Example 23

Alpha-cyano-3,4-dimethoxy-4'-bromostilbene (yellow crystalline substance), 3.17 g (yield: 92.1%), was obtained by the same procedure as in Example 2 except that 1.85 g (10.0 mmol) of 4-bromobenzaldehyde (purchased from Tokyo Kasei) was used instead of 4-chlorobenzaldehyde.

The white crystalline compound represented by the following formula, 544 mg (yield: 86.0%), was obtained by the same procedure as in Example 2 except that 688 mg (2.0 mmol) of the above alpha- Cyano-3,4-dimethoxy-4'-bromostilbene was used instead of alphacyano-3,4-dimethoxy-4'-chlorostilbene: Formula 57:

The melting point of the obtained compound was 171.0ºC ... 171.5ºC.

Example 24

Alpha-cyano-3,4-dimethoxy-4'-methylstilbene (yellow crystalline substance), 2.20 g (yield: 78.8%), was obtained by the same procedure as in Example 2 except that 1.20 g (10.0 mmol) of 4-methylbenzaldehyde (purchased from Wako Junyaku Kogyo) was used instead of 4-chlorobenzaldehyde.

The white crystalline compound represented by the following formula, 438 mg (yield: 87.1%), was obtained by the same procedure as in the example except that 559 mg (2.0 mmol) of the above-mentioned alpha-cyano-3,4-dimethoxy-4'-methylstilbene was used instead of alpha-cyano-3,4-dimethoxy-4'-chlorostilbene: Formula 58:

The melting point of the obtained compound was 162.5ºC ... 163.0ºC.

Example 25

Alpha-cyano-3,4-dimethoxy-3'-methylstilbene (yellow crystalline substance), 1.46 g (yield: 52.3%), was obtained by the same procedure as in Example 2 except that 1.20 g (10.0 mmol) of 3-methylbenzaldehyde (purchased from Tokyo Kasei) was used instead of 4-chlorobenzaldehyde.

The yellow crystalline compound represented by the following formula, 294 mg (yield: 58.5%), was obtained by the same procedure as in Example 2 except that 559 mg (2.0 mmol) of the above-mentioned alpha-cyano-3,4-dimethoxy-3'-methylstilbene was used instead of alpha-cyano-3,4-dimethoxy-4'-chlorostilbene: Formula 59:

The melting point of the obtained compound was 110ºC ... 111ºC.

Example 26

Alpha-cyano-3,4-dimethoxy-4'-ethylstilbene (light yellow crystalline substance), 1.79 g (yield: 61.0%), was obtained by the same procedure as in Example 2 except that 1.34 g (10.0 mmol) of 4-ethylbenzaldehyde (purchased from Tokyo Kasei) was used instead of 4-chlorobenzaldehyde.

The yellow crystalline compound represented by the following formula, 448 mg (yield: 84.4%), was obtained by the same procedure as in Example 2 except that 587 mg (2.0 mmol) of the above-mentioned alpha-cyano-3,4-dimethoxy-4'-ethylstilbene was used instead of alpha-cyano-3,4-dimethoxy-4'-chlorostilbene: Formula 60:

The melting point of the obtained compound was 140.5ºC ... 141.0ºC.

Example 27

The alpha-(3,4-dimethoxyphenyl)-beta-(5-methylthiophen-2-yl)acrylonitrile (light yellow crystalline substance), 2.36 g (yield: 82.7%), was obtained by the same procedure as in Example 2 except that 1.26 g (10.0 mmol) of 5-methylthiophene-2-carboxaldehyde (purchased from Tokyo Kasei) was used instead of 4-chlorobenzaldehyde.

The orange crystalline compound represented by the following formula, 400 mg (yield: 77.7%), was obtained by the same procedure as in Example 2 except that 571 mg (2.0 mmol) of the above-mentioned alpha-(3,4-dimethoxyphenyl)-beta-(5-methylthiophen-2-yl)acrylonitrile was used instead of alpha-cyano-3,4-dimethoxy-4'-chlorostilbene: Formula 61:

The melting point of the obtained compound was 183.0ºC ... 183.5ºC.

Example 28

Alpha-(3,4-dimethoxyphenyl)-beta-(5-methylfuran-2-yl)acrylonitrile (yellow crystalline substance), 2.16 g (yield: 80.2%) was obtained by the same procedure as in Example 2 except that 1.10 g (10.0 mmol) of 5-methylfuran-2-carboxaldehyde (purchased from Wako Junyako Kogyo) was used instead of 3-chlorobenzaldehyde.

The orange crystalline compound represented by the following formula, 320 mg (yield: 66.3%), was obtained by the same procedure as in Example 2 except that 539 mg (2.0 mmol) of the above-mentioned alpha(3,4-dimethoxyphenyl)-beta-(5-methylfuran-2-yl)acrylonitrile was used instead of alpha-cyano-3,4-dimethoxy-4'-chlorostilbene: Formula 62:

The melting point of the obtained compound was 150.5ºC ... 151.0ºC.

Example 29

A mixture of 5-methylfuran-2-carboxaldehyde (3.51 g, 50 mmol, purchased from Wako Junyako Kogyo), rhodanine (6.66 g, 50 mmol, purchased from Tokyo Kasei), and anhydrous sodium acetate (2.3 g, 150 mmol, purchased from Kokusan Kagaku) was refluxed in 35 mL of acetic acid (purchased from Wako Junyako Kogyo) for 30 minutes. After cooling, the reaction mixture was poured into 500 mL of water. The precipitate was collected and washed with water, ethanol, and then ether to obtain 10.04 g (yield: 89.1%) of (5-methylfurylidene)rhodanine. to give an orange-brown colored crystalline substance.

A suspension of the above compound was heated in 65 ml of 15% sodium hydroxide solution at 100°C for 30 minutes. After cooling, the reaction mixture was poured into 500 ml of 10% hydrochloric acid. The precipitate was collected and washed with water to give 8.22 g of 3-(5-methylfuryl)-2-thioketopropanoic acid as a yellow crystalline substance.

To a mixture of 8.22 g of the above compound, 45 ml of ethanol and hydroxylamine hydrochloride (10.1 g, 146 mmol, purchased from Wako Junyaku Kogyo), 55 ml of sodium ethoxide in 21% denatured alcohol solution (147 mmol equivalent, purchased from Aldrich) was gradually added and heated at 100 °C. After cooling, the mixture was concentrated under reduced pressure. To the residue was added 20 ml of 5% sodium hydroxide solution and then 20 ml of 10% hydrochloric acid was carefully added under ice cooling. The mixture was extracted with ether three times and the combined organic layer was washed with brine and dried over anhydrous sodium sulfate. The solvent was removed. The residue was dissolved in a small amount of ether, and then toluene was added to the solution until the mixture was slightly turbid, after which the mixture was allowed to stand. The precipitate was collected and washed with toluene to give 4.55 g (yield: 53.3%) of 3-(5-methyl-2-furyl)-2-hydroxyiminopropanoic acid as a yellow crystalline substance.

To a solution of 4.55 g of the above compound in 60 mL of benzene, 1,1'-carbonyldiimidazole (3.97 g, 24.8 mmol, purchased from Aldrich) was gradually added and the mixture was heated at 70°C for 1 hour. After cooling, the mixture was poured into 50 mL of ice water and extracted three times with benzene. The combined organic layer was washed with aqueous sodium hydrogen carbonate, with brine, with 1% hydrochloric acid and then with water in this order and dried over anhydrous sodium sulfite. The solvent was removed under The residual oil (2.4 g) was purified by chromatography on silica gel (ethyl acetate:hexane = 1:4) to give 2.00 g (yield: 66.6%) of 5-methylfuran-2-yl-acetonitrile as a colorless oily substance.

5-Methyl-alpha-(3,4-dimethoxybenzylidene)- furan-2-acetonitrile (yellow crystals), 419 mg (yield: 31.1%), was obtained by the same procedure as in Example 2 except that 606 mg (5.0 mmol) of the above-mentioned 5-methylfuran-2-yl-acetonitrile and 831 mg (5.0 mmol) of 3,4-dimethoxybenzaldehyde (purchased from Tokyo Kasei) were used.

The orange crystalline compound represented by the following formula, 140 mg (yield: 48.4%), was obtained by the same procedure as in Example 2 except that 323 mg (1.2 mmol) of the above-mentioned 5-methyl-alpha-(3,4-dimethoxybenzylidene)furan-2-acetonitrile and 1.73 g (15 mmol) of pyridinium chloride (purchased from Wako Junyaku Kogyo) were used instead of alpha-cyano-3,4-dimethoxy-4'-chlorostilbene: Formula 63:

The melting point of the obtained compound was 153.0ºC ... 153.5ºC.

The measured values of nuclear magnetic resonance spectra and infrared absorption spectra of the compounds and intermediates of the third embodiment of the present invention prepared in the following Reference Examples and Examples are shown in Tables 14 to 22. The nuclear magnetic resonance spectra [1H-NMR (500 MHz)] were measured in a DMSO-d6 solution with tetramethylsilane as an internal standard or in a mixed solution of CDCl3 with a few drops of CD3OD added; the IR spectra were measured by a KBr tablet method. Table 14 Table 15 Table 16 Table 17 Table 18 Table 19 Table 20 Table 21 Table 22

(Reference example 1) [Preparation of alpha-cyano-4-chloro-3',4'-dihydrostilbene)

To a mixture of 758 mg (5.00 mmol) of 4-chlorophenylacetonitrile (purchased from Tokyo Kasei), 691 mg (5.00 mmol) of 3,4-dihydroxybenzaldehyde (purchased from Tokyo Kasei) and 10 mL of ethanol (purchased from Kokusan Kagaku) was added 0.54 mL of piperidine (purchased from Wako Junyaku Kogyo), the mixture was refluxed with heating for 6 hours and air-cooled, the reaction product was added to 100 mL of 1 N hydrochloric acid, and the precipitates were filtered, washed with water, dissolved in ethyl acetate, washed with a 20% sodium hydrogen sulfite solution twice and then washed with saturated saline once and dried over anhydrous sodium sulfate. Then, the solvent was distilled off under reduced pressure and the residue was purified by column chromatography on silica gel (effluent with a ratio of hexane/ethyl acetate of 5:1 to 5:2) to obtain 919 mg (yield: 67.6%) of alpha-cyano-4-chloro-3',4'-dihydroxystilbene (yellow crystals) represented by the following formula: Formula 32b:

(Reference example 2) [Preparation of alpha-cyano-3'-chloro-3,4-dimethoxystilbene]

3,4-Dimethoxyphenylacetonitrile (purchased from Tokyo Kasei), 1.77 g (10.0 mmol) and 1.41 g (10.0 mmol) of 3-chlorobenzaldehyde (purchased from Tokyo Kasei) were dissolved in 10 ml of ethanol (purchased from Kokusan Kagaku) with heating, two drops of a 20% sodium hydroxide solution were added, and the mixture was stirred overnight. and the precipitated crystals were filtered, washed with ethanol twice and then washed with hexane twice and dried to obtain 2.35 g (yield: 78.3%) of alpha-cyano-3'-chloro-3,4-dimethoxystilbene (yellow crystals).

(Reference example 3) [Preparation of alpha-cyano-3'-chloro-3,4-hydroxystilbene]

A mixture of 600 mg (2.00 mmol) of alpha-cyano-3'-chloro-3,4-dimethoxystilbene obtained by the same procedure as in Reference Example 2 and 3.5 g (30 mmol) of pyridinium chloride (purchased from Wako Junyaku Togyo) was melted and mixed on an oil bath previously heated to 210 °C in an argon atmosphere, stirred at the same temperature for 1 hour and air-cooled, 20 ml of 2N hydrochloric acid was added to the solidified reaction product, the mixture was crushed and stirred for 30 minutes, and the precipitates were then filtered, washed three times with water, dried and purified by means of a short silica gel column (ethyl acetate was used as the effluent) to obtain 436 mg (yield: 80.2%) of alpha-cyano-3'-chloro-3,4-dihydroxystilbene (yellow crystals): Formula 33b:

(Reference example 4) [Production of 4-iodophenylacetonitrile]

A mixture of p-iodotoluene (4.36 g, 20 mmol, purchased from Tokyo Kasei) and N-bromosuccinimide (3.92 g, 22 mmol, purchased from Tokyo Kasei) in 60 ml of carbon tetrachloride (purchased from Wako Junyaku Kogyo) was refluxed for 4 hours under irradiation with an incandescent lamp, to give 2.67 g (yield: 45.0%) of 4-iodobenzyl bromide as a white crystalline substance.

To a hot solution (50 °C) of sodium cyanide (0.49 g, 10.0 mmol, purchased from Kokusan Kagaku) in 10 mL of dimethyl sulfoxide (purchased from Aldrich), the above-mentioned 4-iodobenzyl bromide (1.48 g, 5.0 mmol) was added, and the mixture was stirred for 3 hours at ambient temperature. To the resulting reaction mass was added water and extracted with hexane. The solvent was removed to give 0.84 g (yield: 69%) of 4-iodophenylacetonitrile as a white crystalline substance.

(Reference example 5) [Preparation of 4-bromothiophen-2-yl-acetonitrile]

To a solution of 4-bromothiophene-2-carboxaldehyde (9.55 g, 50 mmol, purchased from Aldrich) in 100 mL of ethanol was added sodium borohydride (3.78 g, 100 mmol, purchased from Yoneyama Yakuhin) gradually with cooling in an ice bath. After the addition was completed, the mixture was stirred at room temperature for an additional 1.5 hours. The reaction mixture was acidified with hydrochloric acid and then concentrated to dryness under reduced pressure. Water was added to the residue and it was extracted with ether to give 9.29 g (yield: 96%) of 4-bromothiophene-2-yl-methanol as an oil.

A mixture of the above-mentioned 4-bromothiophen-2-yl-methanol (4.83 g, 25 mmol) and 63 ml of 47% hydrobromic acid was vigorously stirred for 30 minutes at room temperature. The reaction mixture was extracted with pentane to give 5.31 g (yield: 82.8%) of 2-bromomethyl-4-bromothiophene as a pale yellow oil. The 5.12 g of 2-bromomethyl-4-bromothiophene thus obtained was reacted with sodium cyanide according to the same procedure as in the synthesis of 4-iodophenylacetonitrile of Example 5 to give 1.50 g (yield: 37.0%) of 4-bromothiophen-2-yl-acetonitrile as white crystals, after which the by-product alpha,alpha-bis[(4-bromothiophene-2- yl)methyl]-4-bromothiopheneacetonitrile was removed by recrystallization.

Example 30

The alpha-cyano-4-chloro-3',4'-dihydroxystilbene, 136 mg (0.50 mmol) obtained by the same procedure as in Reference Example 1 was suspended in 1 ml of acetic anhydride (purchased from Wako Junyaku Kogyo), 0.5 ml of triethylamine (purchased from Kokusan Kagaku) was added thereto under ice-cooling and stirring, and then stirred at room temperature overnight, the obtained reaction product was added to 15 ml of 1 N hydrochloric acid and stirred for 30 minutes, and the precipitated crystals were filtered, washed with water, dried and recrystallized from ethyl acetate/hexane to obtain 147 mg (yield: 82.6%) as a white crystalline compound represented by the following formula: Formula 34b:

The melting point of the obtained compound was 119.5ºC ... 120ºC.

Example 31

The alpha-cyano-4-chloro-3',4'-dihydroxystilbene, 543 mg (2.00 mmol), obtained by the same procedure as in Reference Example 1, was suspended in 10 ml of methylene chloride (purchased from Kokusan Kagaku), 0.69 ml of triethylamine (purchased from Kokusan Kagaku) was added and stirred, after which the obtained orange transparent solution was ice-cooled, 0.44 ml of propionyl chloride (purchased from Tokyo Kasei) was added dropwise and stirred under ice-cooling, this was then stirred at room temperature for 1 hour, ethyl acetate was added to the obtained reaction product, which resulting product was washed with 1 N hydrochloric acid, a sodium hydrogen carbonate solution and a saturated saline solution in this order and dried over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure and the residue was recrystallized from ethyl acetate/hexane to obtain 664 mg (yield: 86.5%) as a white crystalline compound represented by the following formula: Formula 35b:

The melting point of the obtained compound was 107.0ºC ... 107.5ºC.

Example 32

The white crystalline compound represented by the following formula, 740 mg (yield: 89.8%), was obtained by the same procedure as in Example 31 except that 0.52 ml of butyryl chloride (purchased from Tokyo Kasei) was used instead of propionyl chloride: Formula 36b:

The melting point of the obtained compound was 86.0ºC ... 86.5ºC.

Example 33

The white crystalline compound represented by the following formula, 428 mg (yield: 45.7%), was obtained by the same procedure as in Example 31 except that 0.69 ml of hexanoyl chloride (purchased from Tokyo Kasei) was used instead of propionyl chloride: Formula 37b:

The melting point of the obtained compound was 61.5ºC ... 62.0ºC.

Example 34

Alpha-cyano-4'-chloro-3,4-dimethoxystilbene, 2.39 g (yield: 79.7%), was obtained by the same procedure as in Reference Example 2 except that 1.41 g (10 mmol) of 4-chlorobenzaldehyde (purchased from Tokyo Kasei) was used instead of 3-chlorobenzaldehyde.

Alpha-cyano-4'-chloro-3,4-dihydroxystilbene, 455 mg (yield: 67.0%), represented by the following formula was obtained by the same procedure as in Reference Example 3 except that 749 mg (2.50 mmol) of the above-mentioned alpha-cyano-4'-chloro-3,4-dimethoxystilbene was used instead of alpha-cyano-3'-chloro-3,4-dimethoxystilbene and that 2.3 g (20 mmol) of pyridinium (purchased from Wako Junyaku Kogyo) was used: Formula 38b:

The white crystalline compound represented by the following formula, 153 mg (yield: 86.0%), was obtained by the same procedure as in Example 30 except that 136 mg (0.50 mmol) of the above-mentioned alpha-cyano-4'-chloro-3,4-dihydroxystilbene was used instead of alpha-cyano-4-chloro-3',4'-dihydroxystilbene. Formula 39b:

The melting point of the obtained compound was 130.0ºC ... 130.5ºC.

Example 35

The white crystalline compound represented by the following formula, 109 mg (yield: 61.5%), was obtained by the same procedure as in Example 30 except that 136 mg (0.50 mmol) of alpha-cyano-3'-chloro-3,4-dihydroxystilbene obtained by the same procedure as in Reference Example 3 was used instead of alpha-cyano-4-chloro-3',4'-dihydroxystilbene: Formula 40b:

The melting point of the obtained compound was 114.0ºC ... 114.5ºC.

Example 36

The alpha-cyano-4'-fluoro-3,4-dimethoxystilbene, 2.33 g (yield: 82.2 g), was obtained by the same procedure as in Reference Example 2 except that 1.24 g (10 mmol) of 4-fluorobenzaldehyde (purchased from Tokyo Kasei) was used instead of 3-chlorobenzaldehyde.

Alpha-cyano-4'-fluoro-3,4-dihydroxystilbene, 432 mg (yield: 84.6%), represented by the following formula was obtained by the same procedure as in Reference Example 3 except that 567 g (2.00 mmol) of the above-mentioned alpha-cyano-4'-fluoro-3,4-dimethoxystilbene was used instead of alpha-cyano-3'-chloro-3,4-dimethoxystilbene: Formula 41b:

The white crystalline compound represented by the following formula, 151 mg (yield: 88.8%), was obtained by the same procedure as in Example 30 except that 128 mg (0.50 mmol) of the above-mentioned alpha-cyano-4'-fluoro-3,4-dihydroxystilbene was used instead of alpha-cyano-4-chloro-3',4'-dihydroxystilbene: Formula 42b:

The melting point of the obtained compound was 145.5ºC ... 146.0ºC.

Example 37

Alpha-cyano-4'-bromo-3,4-dimethoxystilbene, 3.17 g (yield: 92.1%), was obtained by the same procedure as in Reference Example 2 except that 1.85 g (10 mmol) of 4-bromobenzaldehyde (purchased from Tokyo Kasei) was used instead of 3-chlorobenzaldehyde.

Alpha-cyano-4'-bromo-3,4-dihydroxystilbene, 544 mg (yield: 86.0%), represented by the following formula was obtained by the same procedure as in Reference Example 3 except that 688 mg (2.00 mmol) of the above-mentioned alpha-cyano-4'-bromo-3,4-dimethoxystilbene was used instead of alpha-cyano-3'-chloro-3,4-dimethoxystilbene: Formula 43b:

The white crystalline compound represented by the following formula, 163 mg (yield: 81.3%), was obtained by the same procedure as in Example 30 except that 158 mg (0.50 mmol) of the above-mentioned alpha-cyano-4'-boron-3,4-dihydroxystilbene was used instead of alpha-cyano-4-chloro-3',4'-dihydroxystilbene: Formula 44b:

The melting point of the obtained compound was 124.5ºC ... 125.0ºC.

Example 38

Alpha-cyano-4-iodo-3',4'-dihydroxystilbene represented by the following formula, 1.39 g (yield: 76.4%), was obtained by the same procedure as in Reference Example 1 except that 1.22 g (5.00 mmol) of 4-iodophenylacetonitrile obtained by the same procedure as in Reference Example 4 was used instead of 4-chlorophenylacetonitrile: Formula 45b:

The white crystalline compound represented by the following formula, 187 mg (yield: 83.7%), was obtained by the same procedure as in Example 30 except that 182 mg (0.50 mmol) of the above-mentioned alpha-cyano-4-iodo-3',4'-dihydroxystilbene was used instead of alpha-cyano-4-chloro-3',4'-dihydroxystilbene: Formula 46b:

The melting point of the obtained compound was 156.0ºC ... 156.5ºC.

Example 39

Alpha-cyano-4'-methyl-3,4-dimethoxystilbene, 2.20 g (yield: 78.8%), was obtained by the same procedure as in Reference Example 2 except that 1.20 g (10 mmol) of 4-methylbenzaldehyde (purchased from Wako Junyaku Kogyo) was used instead of 4-chlorobenzaldehyde.

Alpha-cyano-4'-methyl-3,4-dihydroxystilbene, 438 mg (yield: 87.1%), represented by the following formula was obtained by the same procedure as in Reference Example 3 except that 559 mg (2.00 mmol) of the above-mentioned alpha-cyano-4'-methyl-3,4-dimethoxystilbene was used instead of alpha-cyano-3'-chloro-3,4-dimethoxystilbene: Formula 47b:

The white crystalline compound represented by the following formula, 144 mg (yield: 85.7%), was obtained by the same procedure as in Example 30 except that 126 mg (0.50 mmol) of the above-mentioned alpha-cyano-4'-methyl-3,4-dihydroxystilbene was used instead of alpha-cyano-4-chloro-3',4'-dihydroxystilbene: Formula 48b:

The melting point of the obtained compound was 130.5ºC ... 131.0ºC.

Example 40

Alpha-cyano-3'-methyl-3,4-dimethoxystilbene, 1.13 g (yield: 40.5%), was obtained by the same procedure as in Reference Example 2 except that 1.20 g (10.0 mmol) of 3-methylbenzaldehyde (purchased from Wako Junyaku Kogyo) was used instead of 4-chlorobenzaldehyde.

Alpha-cyano-3'-methyl-3,4-dihydroxystilbene represented by the following formula, 294 mg (yield: 58.5%), was obtained by the same procedure as in Reference Example 3 except that 559 mg (2.00 mmol) of the above-mentioned alpha-cyano-3'-methyl-3,4-dimethoxystilbene were used instead of alpha-cyano-3'-chloro-3,4-dimethoxystilbene: Formula 49b:

The white crystalline compound represented by the following formula, 102 mg (yield: 61.0%), was obtained by the same procedure as in Example 30 except that 126 mg (0.50 mmol) of the above-mentioned alpha-cyano-3'-methyl-3,4-dihydroxystilbene was used instead of alpha-cyano-4-chloro-3',4'-dihydroxystilbene. Formula 50b:

The melting point of the obtained compound was 87.5ºC ... 88.0ºC.

Example 41

Alpha-cyano-4'-ethyl-3,4-dimethoxystilbene, 1.79 g (yield: 61.0%), was obtained by the same procedure as in Comparative Example 2 except that 1.34 g (10.0 mmol) of 4-ethylbenzaldehyde (purchased from Tokyo Kasei) was used instead of 3-chlorobenzaldehyde.

Alpha-cyano-4'-ethyl-3,4-dihydroxystilbene represented by the following formula, 448 mg (yield: 84.4%) was obtained by the same procedure as in Reference Example 3 except that 587 mg (2.00 mmol) of the above-mentioned alpha-cyano-4'-ethyl-3,4-dimethoxystilbene was used instead of of alpha-cyano-3'-chloro-3,4-dimethoxystilbene were used: Formula 51b:

The white crystalline compound represented by the following formula, 131 mg (yield: 74.8%), was obtained by the same procedure as in Example 30 except that 133 mg (0.50 mmol) of the above-mentioned alpha-cyano-4'-ethyl-3,4-dihydroxystilbene was used instead of alpha-cyano-4-chloro-3',4'-dihydroxystilbene: Formula 52b:

The melting point of the obtained compound was 90.5ºC ... 91.0ºC.

Example 42

Alpha-cyano-4-trifluoromethyl-3',4'-dihydroxystilbene, 657 mg (yield: 87.4%), represented by the following formula was obtained by the same procedure as in Reference Example 1 except that 926 mg (5.00 mmol) of trifluoromethylphenylacetonitrile (purchased from Adrich) was used instead of 4-chlorophenylacetonitrile: Formula 53b:

The white crystalline compound represented by the following formula, 120 mg (yield: 61.6%), was obtained by the same procedure as in Example 30 except that 153 mg (0.50 mmol) of the above-mentioned alpha-cyano-4-trifluoromethyl-3',4'-dihydroxystilbene was used instead of 4-cyano-4-chloro-3',4'-dihydroxystilbene: Formula 54b:

The melting point of the obtained compound was 138.5ºC ... 139.0 ºC.

Example 43

Alpha-cyano-4-methoxy-3',4'-dihydroxystilbene, 214 mg (yield: 16.0%), represented by the following formula was obtained by the same procedure as in Reference Example 1 except that 736 mg (5.00 mmol) of 4-methoxyphenylacetonitrile (purchased from Aldrich) was used instead of 4-chlorophenylacetonitrol: Formula 55b:

The light yellow crystalline compound represented by the following formula, 87 mg (yield: 50%), was obtained by the same procedure as in Example 30 except that 134 mg (0.50 mmol) of the above-mentioned alpha-cyano-4-methoxy-3',e'-dihydroxystilbene was used instead of alpha-cyano-4-chloro-3',4'-dihydroxystilbene: Formula 56b:

The melting point of the obtained compound was 131.0ºC ... 131.5ºC.

Example 44

Alpha-cyano-3,4-dimethoxystilbene, 2.62 g (yield: 98.9%), was obtained by the same procedure as in Reference Example 2 except that 1.06 g (10.0 mmol) of benzaldehyde (purchased from Wako Junyaku Kogyo) was used instead of 4-chlorobenzaldehyde.

Alpha-cyano-3,4-dihydroxystilbene, 253 mg (yield: 53.4%), represented by the following formula was obtained by the same procedure as in Reference Example 3 except that 531 mg (2.00 mmol) of the above-mentioned alpha-cyano-3,4-dimethoxystilbene was used instead of alpha-cyano-3'-chloro-3,4-dimethoxystilbene: Formula 57b:

The white crystalline compound represented by the following formula, 113 mg (yield: 70.2%), was obtained by the same procedure as in Example 30 except that 119 mg (0.50 mmol) of the above-mentioned alpha-cyano-3,4-dihydroxystilbene was used instead of alpha-cyano-4-chloro-3',4'-dihydroxystilbene: Formula 58b:

The melting point of the obtained compound was 148.5ºC ... 149.0ºC.

Example 45

Alpha-cyano-4-nitro-3',4'-dihydroxystilbene · 1/4 hydrate, 1.18 g (yield: 82.3%), represented by the following formula was obtained by the same procedure as in Reference Example 1 except that 811 mg (5.00 mmol) of 4-nitrophenylacetonitrile (purchased from Aldrich) was used instead of 4-chlorophenylacetonitrile, the reaction product was stirred for 4 hours at room temperature and purification was carried out by recrystallization from ethanol/water: Formula 59b:

The yellow crystalline compound represented by the following formula, 110 mg (yield: 60.2%) was obtained by the same procedure as in Example 31 except that 144 mg (0.50 mmol) of the above-mentioned alpha-cyano-4-nitro-3',4'-dihydroxystilbene · 1/4 hydrate was used instead of alpha-cyano-4-chloro-3',4'-dihydroxystilbene using 0.11 ml of acetyl chloride (purchased from Kokusan Kagaku) instead of propionyl chloride and using 0.21 ml of triethylamine (purchased from Kokusan Kagaku): Formula 60b:

The melting point of the obtained compound was 173.5ºC ... 174.0ºC.

Example 46

The alpha-(thiophen-2-yl-methylidene)-3,4-dimethoxybenzeneacetonitrile, 2.49 g (yield: 91.9%), was obtained by the same procedure as in Reference Example 2 except that 112 g (10.0 mmol) of thiophene-2-carboxaldehyde (purchased from Tokyo Kasei) was used instead of 3-chlorobenzaldehyde.

Alpha-(thiophen-2-yl-methylidene)-3,4-dihydroxybenzeneacetonitrile, 344 mg (yield: 70.7%), represented by the following formula was obtained by the same procedure as in Reference Example 3 except that 543 mg (2.00 mmol) of the above-mentioned alpha-(thiophen-2-yl-methylidene)-3,4-dimethoxybenzeneacetonitrile was used instead of alpha-cyano-3'-chloro-3,4-dimethoxystilbene: Formula 61b:

The light brown crystalline compound represented by the following formula, 122 mg (yield: 74.4%), was obtained by the same procedure as in Example 30 except that 122 mg (0.50 mmol) of the above-mentioned alpha-(thiophen-2-yl-methylidene)-3,4-dihydroxybenzeneacetonitrile was used instead of alpha-cyano-4-chloro-3',4'-dihydroxystilbene: Formula 62b:

The melting point of the obtained compound was 162.0ºC ... 162.5ºC.

Example 47

The alpha-(thiophen-3-yl-methylidene)-3,4-dimethoxybenzeneacetonitrile, 2.12 g (yield: 78.2%), was obtained by the same procedure as in Reference Example 2 except that 1.12 g (10.0 mmol) of thiophene-3-carboxaldehyde (purchased from Tokyo Kasei) was used instead of 3-chlorobenzaldehyde.

Alpha-(thiophen-2-yl-methylidene)-3,4-dihydroxybenzeneacetonitrile, 344 mg (yield: 70.7%) represented by the following formula was obtained by the same procedure as in Reference Example 3 except that 543 mg (2.00 mmol) of the above-mentioned alpha-(thiophen-3-yl-methylidene)-3,4-dimethoxybenzeneacetonitrile was used instead of alpha-cyano-3'-chloro-3,4-dimethoxystilbene: Formula 63b:

The light brown crystalline compound represented by the following formula, 121 mg (yield: 74.2%), was obtained by the same procedure as in Example 30 except that 122 mg (0.50 mmol) of the above-mentioned alpha-(thiophen-3-yl-methylidene)-3,4-dihydroxybenzeneacetonitrile was used instead of alpha-cyano-4-chloro-3',4'-dihydroxystilbene: Formula 64b:

The melting point of the obtained compound was 143.5ºC 144.0ºC.

Example 48

Alpha-(3,4-dihydroxybenzylidene)-4-bromothiophene-2-acetonitrile, 105 g (yield: 65.0%), represented by the following formula was obtained by the same procedure as in Reference Example 1 except that 1.01 g (5.00 mmol) of 4-bromothiophen-2-yl-acetonitrile obtained by the same procedure as in Reference Example 5 was used instead of 4-chlorophenylacetonitrile: Formula 65b:

The light yellow compound represented by the following formula, 150 mg (yield: 73.8%), was obtained after the same procedure as in Example 31 except that 161 mg (0.50 mmol) of the above-mentioned alpha-(3,4-dihydroxybenzylidene)-4-bromothiophene-2-acetonitrile was used instead of alpha-cyano-4-chloro-3',4'-dihydroxystilbene, 0.11 ml of acetyl chloride (purchased from Wako Junyaku Kogyo) was used instead of propionyl chloride, and 0.21 ml of triethylamine (purchased from Kokusan Kagaku) was used: Formula 66b:

The melting point of the obtained compound was 125.5ºC ... 126.0ºC.

Example 49

The alpha-(5-methylthiophen-2-yl-methylidene)-3,4-dimethoxybenzeneacetonitrile, 2.36 g (yield: 82.8%), was obtained by the same procedure as in Reference Example 2 except that 1.26 g (10.0 mmol) of 5-methylthiophene-2-carboxaldehyde (purchased from Tokyo Kasei) was used instead of 3-chlorobenzaldehyde.

Alpha-(5-methylthiophen-2-yl-methylidene)-3,4-dihydroxybenzeneacetonitrile, 400 mg (yield: 77.8%), represented by the following formula was obtained by the same procedure as in Reference Example 3 except that 571 mg (2.00 mmol) of the above-mentioned alpha-(5-methylthiophen-2-yl-methylidene)-3,4-dimethoxybenzeneacetonitrile was used instead of alpha-cyano-3'-chloro-3,4-dimethoxystilbene: Formula 67b:

The orange compound represented by the following formula, 141 m (yield: 82.8%), was obtained by the same procedure as in Example 30 except that 129 mg (0.50 mmol) of the above-mentioned alpha-(5-methylthiophene-2-yl-methylidene)-3,4-dihydroxybenzeneacetonitrile was used instead of alpha-cyano-4-chloro-3',4'-dihydroxystilbene: Formula 68b:

The melting point of the obtained compound was 161.0ºC ... 161.5ºC.

Example 50

The alpha-(5-methylfuran-2-yl-methylidene)-3,4-dimethoxybenzeneacetonitrile, 2.16 g (yield: 80.3%), was obtained by the same procedure as in Reference Example 2 except that 1.10 g (10.0 mmol) of 5-methylfuran-2-carboxaldehyde (purchased from Wako Junyaku Kogyo) was used instead of 3-chlorobenzaldehyde.

Alpha-(5-methylfuran-2-yl-methylidene)-3,4-dihydroxybenzeneacetonitrile represented by the following formula, 320 g (yield: 66.3%), was obtained by the same procedure as in Reference Example 3 except that 539 mg (2.00 mmol) of the above-mentioned alpha-(5-methylfuran-2-yl-methylidene)-3,4-dimethoxybenzeneacetonitrile trils were used instead of alpha-cyano-3'-chloro-3,4-dimethoxystilbene: Formula 69b:

The yellow compound represented by the following formula, 31 mg (yield: 19%), was obtained by the same procedure as in Example 30 except that 121 mg (0.50 mmol) of the above-mentioned alpha-(5-methylfuran-2-yl-methoylidene)-3,4-dihydroxybenzeneacetonitrile was used instead of alpha-cyano-4-chloro-3',4'-dihydroxystilbene: Formula 70b:

The melting point of the obtained compound was 199.5ºC ... 200.0ºC.

Example 51

Alpha-cyano-4-chloro-3',4',5'-trihydroxystilbene, 344 mg (yield: 23.9%), represented by the following formula was obtained by the same procedure as in Reference Example 1 except that 771 mg (5.00 mmol) of 3,4,5-trihydroxybenzaldehyde (purchased from Aldrich) was used instead of 3,4-dihydroxybenzaldehyde: Formula 71b:

The white compound represented by the following formula, 191 mg (yield: 92.4%), was obtained by the same procedure as in Example 31 except that 144 mg (0.50 mmol) of the above-mentioned alpha-cyano-4-chloro-3',4',5'-trihydroxystilbene was used instead of alpha-cyano-4-chloro-3',4'-dihydroxystilbene, 0.14 ml of acetyl chloride (purchased from Wako Junyako Kogyo) was used instead of propionyl chloride, and 0.28 ml of triethylamine (purchased from Kokusa Kagaku) was used: Formula 72b:

The melting point of the obtained compound was 203.0 ... 203.5 °C.

Example 52

A mixture of the following composition was prepared per tablet and prepared using a tablet press according to a conventional procedure to prepare a drug composition which selectively inhibits 12-lipoxygenase according to the present invention:

Compound obtained in Example 34: 20.0 (mg)

Lactose (purchased from Iwaki Saiyaku) 40.0

Grain starch (purchased from Yoshida Seiyaku) 15.0

Magnesium stearate (purchased from Taihei Kagaku) 0.4

Carboxymethylcellulose calcium (purchased from Nichirin Kagaku Kogyo) 20.0

Example 53

A mixture having the following composition per capsule was prepared and placed in a gelatin capsule according to a conventional procedure to produce a pharmaceutical composition that selectively inhibits 12-lipoxygenase according to the present invention:

Compound obtained in Example 34: 20.0 (mg)

Lactose (purchased from Iwaki Saiyaku) 40.0

Finely powdered cellulose (purchased from Nippon Soda) 30.0

Magnesium stearate (purchased from Taihei Kagaku) 3.0

The measured values of nuclear magnetic resonance spectra and infrared absorption spectra of the compounds prepared in the following examples are shown in Table 24. The nuclear magnetic resonance spectra [1H-NMR (500 MHz)] were measured in a DMSO-d6 solvent with tetramethylsilane as an internal standard and the infrared absorption spectra were measured by a KBr tablet method. Table 24

(Reference Example 6) [Preparation of alpha-(3,4-dimethoxyphenylbeta-(naphthalen-2-yl)acrylonitrile]

A mixture of 3,4-dimethoxyphenylacetonitrile (1.77 g, 10.0 mmol, purchased from Tokyo Kasei) and 2-naphthoaldehyde (1.56 g, 10.0 mmol, purchased from Tokyo Kasei) in 30 mL of ethanol was heated until dissolved. Thereafter, two drops of a 20% aqueous solution of sodium hydroxide were added and the reaction mixture was stirred overnight. The precipitate was collected and washed with ethanol and then with hexane and dried to give 2.60 g (yield: 82.5%) of alpha-(3,4-dimethoxyphenylbeta-(naphthalen-2-yl)acrylonitrile as a yellow crystalline substance.

Example 54

To a mixture of thionaphthene-3-acetonitrile (866 mg, 5.0 mmol, purchased from Lancaster) and 3,4-dihydroxybenzaldehyde (691 mg, 5.0 mmol, purchased from Tokyo Kasei) in 10 mL of ethanol was added 0.54 mL of piperidine (purchased from Wako Junyaku Kogyo) and refluxed for 8 hours. After cooling, the mixture was poured into 100 mL of 1N hydrochloric acid and stirred. The precipitate was collected and washed with water and then dissolved in ethyl acetate. The solution was washed with a 20% aqueous sodium hydrogen sulfite solution twice and then with brine. The organic layer was dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure. The residue was purified by column chromatography on silica gel. Elution with hexane/ethyl acetate (10:1 .. 10:6) gave 580 mg (yield: 39.5%) of the yellow crystalline product represented by the following formula. Formula 22c:

Example 55

The yellow crystalline compound represented by the following formula, 499 mg (yield: 34.7%), was obtained by the same procedure as in Example 54 except that 836 mg (5.00 mmol) of naphthalene-2-acetonitrile (purchased from Aldrich) was used instead of thianaphthene-3-acetonitrile: Formula 23c:

The melting point of the obtained compound was 206ºC ... 207ºC.

Example 56

A mixture of alpha-(3,4-dimethoxyphenylbeta-(naphthalen-2-yl)acrylonitrile (631 mg, 2.0 mmol) was obtained by the same procedure as in Reference Example 6 and melted on an oil bath (210 °C) under argon atmosphere to mix pyridinium chloride (3.5 g, 30 mmol, purchased from Wako Junyaku Kogyo). Stirring was continued for another hour. After cooling, 20 ml of 2N hydrochloric acid was added to the resulting reaction mass and the mixture was stirred for 30 minutes. The precipitate was collected and washed with water three times and then subjected to short column chromatography on silica gel under Elution with ethyl acetate gave 490 mg (yield: 85.3%) of the yellow crystalline product represented by the following formula with mp 166.0ºC ... 166.5ºC: Formula 24c:

Example 57

The yellow crystals of alpha-(3,4-dimethoxyphenyl)- beta -(naphthalen-1-yl)acrylonitrile, 2.42 g (yield: 76.8%), were obtained by the same procedure as in the Reference Examples except that 1.56 g (10.0 mmol) of 1-naphthoaldehyde (purchased from Aldrich) was used instead of 2-naphthoaldehyde.

The yellow crystalline compound represented by the following formula, 524 mg (yield: 91.1%) represented by the following formula 25c was obtained by the same procedure as in Example 56 except that 631 mg (2.00 mmol) of the above-mentioned alpha-(3,4-dimethoxyphenyl)-beta-(naphthalen-1-yl)acrylonitrile was used instead of alpha-(3,4-dimethoxyphenyl)-beta-(naphthalen-2-yl)acrylonitrile: Formula 25c:

The melting point of the obtained compound was 162ºC ... 163ºC.

Example 58

The yellow crystals of alpha-(3,4-dimethoxyphenyl)-beta-(benzofuran-2-yl)acrylonitrile, 2.46 g (yield: 80.7%), were obtained by the same procedure as in Reference Examples except that 1.46 g (10.0 mmol) of benzofuran-2-carboxaldehyde (purchased from Lancaster) was used instead of 2-naphthoaldehyde.

The yellow crystalline compound represented by the following formula, 457 ml (yield: 82.4%), was obtained by the same procedure as in Example 56 except that 611 mg (2.00 mmol) of the above-mentioned alpha-(3,4-dimethoxyphenyl)-beta-(benzofuran-2-yl)acrylonitrile was used instead of alpha-(3,4-dimethoxyphenylbeta-(naphthalen-2-yl)acrylonitrile:

The melting point of the obtained compound was 192ºC ... 193ºC.

Example 59

A mixture of the following composition was prepared per tablet and tabletted using a tablet press in the usual manner to produce a pharmaceutical composition which selectively inhibits 12-lipoxygenase according to the present invention:

Compound obtained in Example 58: 20.0 (mg)

Lactose (purchased from Iwaki Saiyaku) 40.0

Grain starch (purchased from Yoshida Seiyaku) 15.0

Magnesium stearate (purchased from Taihei Kagaku) 0.4

Carboxymethylcellulose calcium (purchased from Nichirin Kagaku Kogyo) 20.0

Example 60

A mixture of the following composition was prepared per capsule and placed in a gelatin capsule according to a conventional procedure to produce a pharmaceutical composition according to the present invention which selectively inhibits 12-lipoxygenase:

Compound obtained in Example 58: 20.0 (mg)

Lactose (purchased from Iwaki Saiyaku) 40.0

Finely powdered cellulose (purchased from Nippon Soda) 30.0

Magnesium stearate (purchased from Taihei Kagaku) 3.0

Possibilities for technical use

As described in detail above, the present invention relates to a stilbene derivative and a stilbene homolog derivative which selectively inhibit 12-lipoxygenase, and pharmaceutical compositions containing the above-mentioned compounds as effective components and which selectively inhibit 12-lipoxygenase; furthermore, the present invention relates to novel compounds of a stilbene derivative and a stilbene homolog derivative which are capable of producing a substance whose modified part is cleaved in vivo and which selectively inhibits 12-lipoxygenase; and pharmaceutical compositions containing the above-mentioned compounds as effective components and which are capable of selectively inhibiting 12-lipoxygenase; and ways of technically utilizing the present invention as follows:

1) The compounds of the present invention have a 12-lipoxygenase inhibitory activity with a strong and high selectivity;

2) the pharmaceutical compositions containing the above-mentioned compounds of the present invention as effective components are useful for the prevention and cure of various circulatory diseases such as arteriosclerosis and vascular spasms, as well as for the prevention of metastasis of some carcinomas;

3) the compounds of the present invention are effective as effective components of drugs for selectively retarding 12-lipoxygenase with low toxicity and few side effects;

4) the compounds of the present invention have a 12-lipoxygenase inhibitory activity with a strong high selectivity since their modified parts are cleaved in vivo by the action of enzymes;

5) it is possible to control the cleavage of the compounds of the invention in vivo by selecting the types of acyl groups as modified parts of the compounds of the invention.

Claims (6)

1. Stilbene derivative or stilbene homologue derivative of the general formula 1: formula 1 wherein R¹ represents a hydrogen atom or a hydroxy group; R² and R³ represent hydrogen atoms or cyano groups (where R² and R³ are different); and Ar is a group represented by the following general formulas 2, 3 or 4: Formula 2 Formula 3 Formula 4 wherein R⁴ represents a hydrogen atom, a methyl group, an ethyl group, a methoxy group, a halogen atom, a trifluoromethyl group or a cyano group. 2. Medicaments containing a compound selected from the group consisting of a stilbene derivative and a stilbene homologue derivative of the following general formula 5: Formula 5 wherein R¹ represents a hydrogen atom or a hydroxy group; R² and R³ represent hydrogen atoms or cyano groups (where R² and R³ are different); and Ar is a group represented by the following general formulas 6, 7 or 8: Formula 6 Formula 7 Formula 8 wherein R⁴ represents a hydrogen atom, a methyl group, an ethyl group, a methoxy group, a halogen atom, a trifluoromethyl group, a cyano group or a nitro group or a mixture thereof as an effective component. 3. Stilbene derivative or stilbene homologue derivative of general formula 1b: Formula 1b wherein R¹ represents a hydrogen atom or OZ, R² and R³ represent hydrogen atoms or cyano groups (where R² and R³ are different); and Ar is a group represented by the following general formulas 2b, 3b or 4b: Formula 2b Formula 3b Formula 4b wherein R⁴ represents a hydrogen atom, a methyl group, an ethyl group, a methoxy group, a halogen atom, a trifluoromethyl group, a cyano group or a nitro group; Z is independently a hydrogen group or a group represented by the following general formula 5b (all Z are not hydrogen atoms): Formula 5b wherein R⁵ represents a straight-chain or branched-chain alkyl group or alkenyl group having 1 to 20 carbon atoms. 4. Medicaments containing a compound selected from the group consisting of a stilbene derivative and a stilbene homologue derivative of the following general formula 6b: Formula 6b wherein R⁴ represents a hydrogen atom or OZ; R² and R³ represent hydrogen atoms or cyano groups (where R² and R³ are different); and Ar is a group, represented by the following general formulas 7b, 8b or 9b: Formula 7b Formula 8b Formula 9b wherein R⁴ represents a hydrogen atom, a methyl group; an ethyl group, a methoxy group, a halogen atom, a trifluoromethyl group, a cyano group or a nitro group; Z is independently a hydrogen group or a group represented by the following general formula 10b (all Z are not hydrogen atoms): Formula 10b wherein R⁵ represents a straight-chain or branched-chain alkyl group or alkenyl group having 1 to 20 carbon atoms or a mixture thereof as an effective component. 5. α,β-Diarylacrylonitrile derivative of the general formula 1c: Formula 1c wherein R¹ represents a hydrogen atom or a hydroxy group; R² and R³ represent hydrogen atoms or cyano groups (where R² and R³ are different); and Ar is a group represented by the following general formulas 2c or 3c: Formula 2c Formula 3c where X represents an oxygen atom or a sulfur atom. 6. Medicaments containing a compound selected from a group consisting of α,β-diarylacrylonitrile derivatives of the general formula 4c: Formula 4c wherein R¹ represents a hydrogen atom or a hydroxy group; R² and R³ represent hydrogen atoms or cyano groups (where R² and R³ are different); and Ar is a group represented by the following general Formulas 5c Formula 6c wherein X represents an oxygen atom or a sulfur atom or a mixture thereof as an effective component.